Enzyme production

Banana peel is one of the wastes from bananas with a composition of 35-40% of the weight of fresh bananas. The use of banana peels as feed and food ingredients is very limited due to the low protein content in banana peels. This study aimed to increase the protein content of banana peel flour using the solid-state fermentation method by Rhizopus oryzae FNCC 6157 in a tray bioreactor. Optimization of protein production of banana peel flour was done using the central composite design by response surface methodology. The variables used in this study were substrate thickness (1 to 3 cm), aeration rate (2 to 4 L/min), and moisture content (50 to 70%). Statistical analysis was performed using ANOVA and resulted in an excellent quadratic equation model as indicated by the F-value and p-value of 17.77 and 0.0012, respectively, and was statistically significant (p≤0.

Bacteria are attracting the interest of worldwide investors; their use is
interesting in several industrial fields, this organism produces a wide variety of extracellular enzymes, including proteases. The objective of this study was to evaluate the production of protease by different bacterial strains isolated from local marine samples collected from the Cap Rousseau beach in the Oran city, northwestern Algeria, 44 bacterial isolates were tested for protease production by cultivating them on skim milk agar medium. The proteolytic activities of all strains were tested using skim milk agar and gelatin agar. Among the 14 isolates that showed a significant hydrolysis diameter, two bacterial strains EC23 and EC2S3 demonstrated the highest potential for protease production and they were selected for further studies. In addition, the extracellular protease was examined using the fermentation production medium at 30°C for 48h, with a constant agitation of 150 rpm. The enzyme activity was determined under varying conditions of pH, incubation temperature, and salt concentration, using Sigma’s
Universal Protease Activity Assay. The enzyme from EC23 strain showed higher activity in all cases than the EC2S3 strain, which indicated that it was the most ideal organism for enzyme production.

The main goal of this study was the characterization of the extremophile xylanases produced by Anoxybacillus sp. strain 3M, a thermophilic bacterium isolated from terrestrial hot springs (temperature of 90°C) samples collected on S. Miguel, Azores, Portugal. In this context, several batch fermentations using different agroindustrial wastes as inducer substrates (BSG -Brewers' spent grain, wheat straw, sugarcane bagasse, and corn cobs) were performed towards an optimal enzyme production. The results for xylanase production showed that the higher enzymatic levels were obtained in the growth medium containing 1% (w/v) BSG (1.34 U mL -1 ), indicating that the BSG was the best inducer substrate, but the xylanase activity was also observed when wheat straw (1.33 U mL -1 ), sugarcane bagasse (0.80 U mL -1 ), corn cobs (0.30 U mL -1 ) and commercial xylan (0.21 U mL -1 ) were used as inducers. The extracellular crude enzymatic extract produced by Anoxybacillus sp. 3M in optimized submerged fermentation with BSG, attaining a maximal xylanase activity of 1.41 U mL -1 (i.e. 5-fold higher than with xylan), was further characterized for its optimal temperature and pH and stability. The highest enzyme activity was observed at a temperature of 60ºC and pH 5.3, but the enzyme retained 100% of its original activity after 96 h at 60°C and pH 7.0 and it still maintained 46% activity after 28 days at the same conditions. At 60ºC, the enzyme also retained its activity for pH ranging from 7.0 to 10.5. In addition, a zymogram of native gel analysis of the different culture supernatants revealed the presence of an enzymatic complex with an apparent molecular weight ranging from 400-500 kDa. Further application of strain 3M enzymatic extract to commercial oat spelts xylan revealed the presence of xylose and xylooligosaccharides (XOS), mainly X 2 and X 3 , in the hydrolyzates produced. So, the thermostable and alkalistable extracellular xylanases from Anoxybacillus sp. 3M are biocatalysts with high potential for the development of processes where lignocellulosics are converted to precursors for several biotechnological applications.

Nigeria is blessed with abundant natural source of renewable energy the; solar, wind, hydro and biomass. However, despite its strategic location near the equatorial region, the use of solar as a source of energy in the country is very insignificant. The current study focused on the spatial analysis of site suitability for solar photovoltaic (PV) power plant in the study area. Administrative map of the study area, Global Horizontal Irradiance (GHI), temperature, elevation, sentinel imagery, power line and road data were used for the study. Analysis was conducted using analytical hierarchical process (AHP) excel template and ArcGIS 10.8 software. Results on the suitability of economic factors revealed that, highly suitable area recorded the highest percentage for aspect (45.4%) and slope (99.1%), while unsuitable area for LULC (67.5%), road (53.1) and power line (59.01) recorded the highest percentage. Results for technical factors showed highly suitable with 66.4% and moderately suitable with 79.9% to be occupying more land area for GHI and temperature respectively. The result on the comparison matrix using AHP revealed GHI with 38.6% with high impact while road with only 4% with the lowest at 4% confidential level. Result on the site suitability index for the PV installation indicated moderately suitable area to has occupied the largest area with 73.7% while unsuitable with the lowest (0.9%) percent. Findings generally indicated that, there are five optimum sites for solar PV plant in the area with the capacity of PV output ranging from 1453.08 to 1459.37 kwh/m 2 /year. Based on these, it is concluded that, there are potential site for solar PV plant in Kaduna metropolis and these sites can provide the amount of energy required. Hence, the study recommends the provision of this type of plant in the area in order to reduce the much pressure on hydro power supply.

Schistosomiasis, a parasitic disease spread by snails, is a major health and economic issue in tropical and subtropical areas, particularly in Pakwach District, Northern Uganda, where it significantly affects both humans and cattle. This study aims to develop a mathematical model to understand the transmission dynamics of schistosomiasis, with a focus on the role of varied intermediate hosts in the region. The model uses nonlinear differential equations and includes human, animal, and snail populations to explore the effectiveness of control measures like mass drug administration, snail control, and public health education. Key findings highlight that controlling the environment where infected snails thrive is crucial, and public health education is especially effective. Limitations include reliance on existing data, and future research should focus on refining the model, vaccine development, and evaluating the costeffectiveness of control strategies. The study provides valuable insights for schistosomiasis control in Pakwach and similar regions globally.

Co-production of thermostable extracellular cellulase and xylanase was investigated using bacterial soil isolate. To evaluate the effect of culture conditions on the c-oproduction of both enzymes by Bacillus subtilis 276NS (GenBank accession number JF801740), a Plackett-Burman fractional factorial design was applied. Among the tested variables, yeast extract, sucrose and incubation time were the most significant variables increased cellulase and xylanase productions. Both of xylan and CMC (Carboxymethyl Cellulose) induced the xylanse enzyme production. A near-optimum medium formulation was obtained which increased the cellulase and xylanase enzymes 5.7-and 1.08fold higher than the yield obtained with the basal medium, respectively. Thereafter, the response surface methodology was adopted to acquire the best process conditions among the selected variables (xylan, CMC and Yeast Extract (YE)) required for improving xylanase yield. The optimal combinations of the major medium constituents for xylanase production evaluated using non-linear optimization algorithm of EXCEL-solver, was as follows (g/L): D-sucrose, 10; xylan, 10.367, CMC, 10.535; (NH4)2 SO4, 1.0; YE, 1.71; Tween-80, 0.4 and FeSO4, 0.25 mg/L, at pH 8.0, temperature 35 C and incubation time 24h under shaking. The predicted optimum thermostable xylanase activity was 360 U/ml, which was around 4-times the activity with the basal medium.

Xylanase-producing bacteria play a crucial role in the degradation of lignocellulosic biomass, facilitating the breakdown of xylan into xylooligosaccharides and xylose. This study aimed to isolate and characterize xylanase-producing bacteria from mangrove ecosystems in Cuddalore and Pondicherry, India. A total of 22 bacterial isolates were obtained, out of which five (PMS1, PMS8, PYMW, SBMS4, and SBW1) exhibited significant xylanolytic activity. Molecular characterization using 16S rRNA sequencing identified these isolates as belonging to the Bacillus genus, with Bacillus aryabhattai (PYMW) and Bacillus subtilis (PMS8) demonstrating the highest xylanase activity. Optimization studies revealed that enzyme production peaked at pH 8.0, with maltose and glucose serving as the most effective carbon sources, and yeast extract enhancing enzyme yield (4.1 ± 0.2 U/mL) and (3.2 ± 0.2 U/mL) respectively. The xylanase enzyme was purified using ammonium sulfate precipitation, yielding a molecular weight of 50 kDa as confirmed by SDS-PAGE. Further, immobilization using sodium alginate beads enhanced enzyme stability and reusability, making it a viable candidate for industrial applications. Molecular docking studies provided insights into enzyme-substrate interactions, highlighting key residues involved in xylan degradation. These findings underscore the potential of mangrove-derived Bacillus strains as sustainable sources of xylanase for applications in bioethanol production, paper manufacturing, and waste management. Future research should focus on large-scale production, genetic modifications for enhanced efficiency, and industrial applications of immobilized xylanase.

Background: Ethanol has been pointed out as a laccase inducer. However, there are controversial reports about its efficiency with some fungi. In this study, we hypothesized that ethanol laccase induced in Pycnoporus sanguineus depends on nitrogen nutriment conditions. To prove this, we assessed laccase production in submerged cultures of P. sanguineus, with different nitrogen concentrations and with, or without ethanol added in a factorial designed experiment. Results: In order to analyze the effects of factors on the response variables, a factorial ANOVA, and response-surface models were performed. It was found that the nitrogen source was the main factor that affected laccase production in P. sanguineus. The treatments with yeast extract (2 g/L) and ethanol (3 g/L) induced the highest laccase activity (31.01 ± 4.9 U/L), while the treatments with urea reached the lowest activity (less than 1.6 U/L). Ethanol had positive and synergic effects on laccase production, in accordance with the surface response model, as long as simple nitrogen sources (urea) were not available. Conclusions: We suggest that laccase in P. sanguineus is regulated by a catabolic nitrogen repression mechanism; laccase activity is strongly inhibited by urea used as nitrogen source and it decreases when the amount of urea increases; contrarily, a synergic positive effect was observed between yeast extract and ethanol on laccase production.

Research Professor, has an extensive expertise in yeast research (Saccharomyces cerevisiae, S. pastorianus, Candida albicans, and C. glabrata) and single-molecule biophysics (high-resolution microscopy, i.e., confocal laser microscopy, AFM, force spectroscopy, and scanning probe lithography), yeast space biology research and hardware development, protein science (yeast adhesins), cell (yeast) immobilization biotechnology, fermentation technology, and brewing science and technology. vii fermentation

Background: Ethanol has been pointed out as a laccase inducer. However, there are controversial reports about its efficiency with some fungi. In this study, we hypothesized that ethanol laccase induced in Pycnoporus sanguineus depends on nitrogen nutriment conditions. To prove this, we assessed laccase production in submerged cultures of P. sanguineus, with different nitrogen concentrations and with, or without ethanol added in a factorial designed experiment. Results: In order to analyze the effects of factors on the response variables, a factorial ANOVA, and response-surface models were performed. It was found that the nitrogen source was the main factor that affected laccase production in P. sanguineus. The treatments with yeast extract (2 g/L) and ethanol (3 g/L) induced the highest laccase activity (31.01 ± 4.9 U/L), while the treatments with urea reached the lowest activity (less than 1.6 U/L). Ethanol had positive and synergic effects on laccase production, in accordance with the surface response model, as long as simple nitrogen sources (urea) were not available. Conclusions: We suggest that laccase in P. sanguineus is regulated by a catabolic nitrogen repression mechanism; laccase activity is strongly inhibited by urea used as nitrogen source and it decreases when the amount of urea increases; contrarily, a synergic positive effect was observed between yeast extract and ethanol on laccase production.

The world’s population is growing at an exponential rate, i.e., 1.59%, and by the end of 2050, the global population has been predicted to grow to an estimated 9 billion people. Furthermore, with the current human population of 7700million and with its rapid growth, food production is expected to increase by no more than 70% by 2050, so feeding the population will be a major challenge. Microbial technology is a type of biotechnology that uses microorganisms in combination with emerging modern biotechnology techniques to develop ecological agriculture. Microscopic technology is concerned with manipulating the genetics of microorganisms or their constituents to make it possible to yield valuable merchandise for a variety of solicitations. In terms of increasing agricultural productivity, traditional farming equipment and practices have reached their limit. Biochemical composts, insect killers, herbicides, and other agricultural inputs have improved agronomic output while lowering soil efficiency and deteriorating conservational eminence. One of the upcoming management strategies is the use of microbial inoculants containing beneficial microbes capable of improving plant growth, quality, and fertility of the soil. Numerous plant-associated contagious groups (nitrogen-fixing, plant growth-promoting rhizobacteria, and mycorrhizal fungi) provide stress resistance by manufacturing a wide variety of enzymes and metabolites. Inoculation of plants with these PGP microbes helps them to bear both biotic and abiotic stresses and promotes plant growth by improving soil structure, moisture retention, mineral absorption, and nutrient uptake. Plants’ genetic dependence on the valuable functions provided by symbiotic cohabitants explains the widespread use of organisms in sustainable agriculture. Biotechnologies that have recently been developed using an ecofriendly and cost-effective approach can provide useful new tools for solving a variety of specific problems in sustainable agriculture. The purpose of this study is to compile a list of various microbial biotechnology applications for sustainable agriculture development.

The increasing demand for energy-efficient and time-optimized computational systems has driven research into combinatorial algorithms, particularly those used to solve the knapsack problem. The knapsack problem is one of the most significant in combinatorial optimization, which involves determining the optimal selection of items to include in a knapsack while adhering to specific constraints, such as weight or profit limits. This study compares the energy consumption and time complexity of the greedy and dynamic programming algorithms applied to this problem. Using power models to measure total energy consumption and execution time, the research reveals that the greedy algorithm is far more efficient, with negligible energy consumption across various scenarios. In contrast, the dynamic programming algorithm, while delivering accurate solutions, consumes more energy and takes longer due to its memory-intensive operations. These findings highlight the need to consider energy and time efficiency in algorithm design, contributing to more sustainable computing practices. Future research will explore larger datasets and focus on instruction-level energy analysis to optimize algorithm performance.

The global fishing industry, a vital source of food and economic activity, generates substantial amounts of fish
scale waste, a byproduct of fish processing that poses significant environmental challenges. This waste, often
discarded in landfills, contributes to environmental pollution and the depletion of valuable resources,
particularly keratin, a protein with diverse industrial applications. Microbial bioconversion of the rich keratinous
waste abundant in the environment provides a rich source of available protein which lurks in keratin-rich waste
and this achievement will lead to a reduction in environmental pollution and improve the economy as the
proteins gotten and microorganisms isolated can be utilized in the production of high-value products in both
agriculture, pharmaceutical, cosmetics, detergent, textile, fertilizer, and biomedical industries. This study aimed
to isolate keratinase-producing organisms from decaying fish scales. The research commenced by enriching
decayed fish scales in a basal medium, followed by inoculation onto three different media: Fish Scale Agar
(FSA), Starch Casein Agar (SCA), and Nutrient Agar (NA). This resulted in the isolation of 27 bacterial strains,
they were subjected to proteolytic enzyme activity analysis on skim milk agar to identify the proteolytic
organisms and six (6) isolates namely SCAA1 (2cm), SCAA2 (1.3cm), SCAA3 (4cm), SCAA4 (4.2cm), FSAA2
(2.4cm) and FSAA5 (2.2cm) showed hallow zones of different magnitude on skim milk agar. FSAA2 (68%) and
SCAA1 (66%) showed highest levels of degradation of whole fish scales and these two isolates were identified
as Alcaligenes faecalis (SCA A1) and Pseudomonas aeruginosa (FSA A2) using a combination of Gram's
reaction, biochemical tests, and 16S rRNA gene sequencing.

Keratinases are enzymes that catalyze the degradation of keratin, a resilient protein found in various biological
materials such as: feathers, hair, and fish scales. Two isolates identified from previous work as Alcaligenes faecalis
(SCA A1) and Pseudomonas aeruginosa (FSA A2) was used for keratinase production. The enzyme produced were
purified using dialysis, ion exchange chromatography and gel filtration chromatography and the purified enzyme was
characterized and used to check for keratin substrate specificity. Fermentation studies revealed that A. faecalis (SCA
A1) displayed optimal growth at pH 9.5 and 10.5, particularly at higher temperatures (48°C, 58°C, and 68°C) when
using Mackerel fish scales as the substrate. In contrast, P. aeruginosa (FSA A2) demonstrated better growth at pH 7.5
and 8.5, with the highest growth observed at 28°C and 38°C using both Mackerel and Elephant fish scales. Enzyme
activity profiles showed that A. faecalis exhibited higher activity with Mackerel scales (65.7 U/ml), while P.
aeruginosa showed greater activity using Elephant scales (68.1 U/ml). Purification of the keratinase enzymes from
both isolates resulted in a 7-fold increase in specific activity, reaching 35.16 U/mg for A. faecalis and 37.80 U/mg for
P. aeruginosa. Characterization of the purified enzymes revealed that both exhibited optimal activity at 70°C, with P.
aeruginosa displaying slightly higher activity. The enzymes also shows good activity over a wide pH range, with
optima at pH 7.3 and 6.3 for A. faecalis and P. aeruginosa, respectively. Substrate specificity analysis indicated that
both enzymes had the highest activity towards Mackerel fish scales, followed by Casein, Human Hair, Chicken
feathers, and Elephant fish scales. These findings highlight the potential of A. faecalis and P. aeruginosa for the
biodegradation of fish scale waste, offering a sustainable solution for waste management and the production of
valuable bioproducts. These results suggest that these keratinolytic bacteria could be utilized in the development of
bioremediation strategies for fish processing waste. Additionally, the keratinase enzymes produced by these isolates
could be utilized in various industrial applications, such as the production of animal feed supplements, leather
processing, and the development of cosmetic and pharmaceutical products.

The enzyme β-galactosidase has two main biotechnological applications in milk and dairy products, e.g. the removal of lactose from milk for lactose-intolerant people and the production of galacto-oligosaccharides (GOS) for use in probiotic food. The intracellular β-galactosidase from thermophile B1.2 was isolated from Ta Pai hot spring, Maehongson, Thailand. The enzyme was purified by ion-exchange and affinity chromatography with a fold purification of 2.2 and 3.9, respectively. The activity of purified β-galactosidase was shown to have an optimal operating condition at pH 6.5 and a temperature of 60 0 C. The thermostability of the enzyme was in the range of 40-60 0 C with the pH stability in the range of 6.0 -10.0. The K m and V max values for oNPG were determined as 28.85 mM and 8.38 ×10 -3 mmolL -1 min -1 , respectively. An intracellular β-galactosidase from B1.2 was also inhibited by various mono-and divalent cations, including Zn + and Mg 2+ ; reagents, including EDTA. It was moderately inhibited by its reaction products; glucose and galactose. The molecular mass of the purified enzyme as determined by native PAGE was approximately 215 kDa, by SDS -PAGE was 75 kDa and by gel filtration was 215 kDa.

The limited hydration capacity and challenges related to delayed expansion prevent fly ash (FA) and MgO expansive additive (MEA) from being used significantly. Nonetheless, utilizing these two procedures in hydraulic mass concrete applications is a frequently used approach that yields favorable outcomes. To construct and assess machine learning-based algorithms to assess the volume expansion (V_e) of cement paste, which consists of FA and MEA, 170 experimental findings from published studies are employed. A novel approach called least square support vector regression (LSSVR) has been developed. The efficacy of LSSVR is significantly impacted by its hyperparameters (c and g), which were fine-tuned using the Dwarf Mongoose Optimization Algorithm (DMOA) and the Equilibrium Optimization Algorithm (EOA). Based on the results obtained, it can be inferred that there exists a significant potential for both 〖LSSVR〗_E and 〖LSSVR〗_D models to accurately predict the V_e of cement paste that incorporates fly ash and MgO expansive addition. In the training and testing phases, the Theil inequality coefficient (TIC) values for 〖LSSVR〗_E are observed to be 0.0906 and 0.01043, which are comparatively higher than the TIC values for 〖LSSVR〗_D, which are 0.0382 and 0.0044, respectively. By predicting the volume expansion accurately, engineers can adjust the proportions of FA and MEA to achieve desired expansion properties, improving the durability and stability of concrete structures. Accurate prediction models allow for better control of thermal stresses, reducing the risk of thermal cracking and extending the structure's lifespan.

The population dynamics of early replicators has revealed numerous puzzles, highlighting the difficulty of transitioning from simple template-directed replicating molecules to complex biological systems. The resolution of these puzzles has set the research agenda on prebiotic evolution since the seminal works of Manfred Eigen in the 1970s. Here, we study the effects of demographic noise on the population dynamics of template-directed (non-enzymatic) and protein-mediated (enzymatic) replicators. We borrow stochastic algorithms from evolutionary game theory to simulate finite populations of two types of replicators. These algorithms recover the replicator equation framework in the infinite population limit. For large but finite populations, we use finite-size scaling to determine the probability of fixation and the mean time to fixation near a threshold that delimits the regions of dominance of each replicator type. Since enzyme-producing replicators cannot evolve in a well-mixed population containing replicators that benefit from the enzyme but do not encode it, we study the evolution of enzyme-producing replicators in a finite population structured in temporarily formed random groups of fixed size n. We argue that this problem is identical to the weak-altruism version of the n-player prisoner’s dilemma, and show that the threshold is given by the condition that the reward for altruistic behavior is equal to its cost.

This paper presents a model-based optimization study for enzyme production in a filamentous fungal fermentation. An existing morphologically structured model is amended by including the effect of low levels of oxygen on growth and production, as well as the effect of medium rheological properties on the oxygen transfer. These changes are needed to extend the applicability of the model to fed-batch operation. Based on this model, optimal feeding strategies are proposed for improving the performance of the process compared to its current operation in industry. The main result is that it is better to reduce the initial substrate concentration (and also the amount of biomass formed), which goes against expected intuition that having more catalyst (biomass) available will improve the performance optimization. Yet, with this strategy, less dead biomass is formed and there is no oxygen limitation problem.

The production of â-galactosidase by Lactobacillus acidophilus NRRL 4495 was studied. The highest yield of intracellular â-galactosidase was achieved using acid whey (3.5%) as carbon source after 48h fermentation. Ammonium sulphate (3%) was the best nitrogen source. Addition of 2 MnCl at 0.02M optimized the production of the enzyme. In addition, the activity reached 51.46U/g cells when the temperature of the fermentation optimized at 40°C. The highest â-galactosidase activity was obtained by using mechanical disintegration of cells by mortar with sterile sea sand under cooling. By studying some properties of the produced enzyme, the enzyme activity was optimum at 40°C and pH 7.0. In absence of substrate and up to 45°C the enzyme was highly stable, at least for 60 min, retaining 83% of its original activity.

During a screening program for antimicrobial compounds from underexplored habitats, a Gram-positive bacterium TD-032, was isolated from arid soil, Thar Desert (India), and analyzed for its morphological, physicochemical, and antimicrobial properties. The 16S ribosomal DNA (rDNA) sequence of the isolate was further studied for the novelty of γ-hyper variable region. TD-032 was grown in large-scale culture, and aqueous and organic solvent extracts analyzed for antimicrobial activity. Culture characteristics showed a lack of diffusible and melanoid pigments. The morphological features were pale yellow aerial mycelium colony color with brownish yellow substrate mycelium and leathery texture. The isolate could grow at 1% concentration of sodium chloride, temperature of 40°C, and a wide range of pH (7.0-12.0). An evaluation for extracellular enzymatic activities showed secretion of gelatinase(s), cellulase(s), and lipase(s). The γ-hyper variable region of 16S rDNA sequence of TD-032 showe...

The mixture of phenol and 4-methylphenol (p-cresol) effect on the growth behavior and degradation capacity of Aspergillus awamori strain was investigated. The phenol hydroxylase activity modulation depending on substrates included in the media was established. The enzyme hydroxylation of benzene-1,4-diol (hydroquinone) or 4-methylbenzene-1,2-diol (p-methylcatechol) goes better compared with phenol or p-cresol substrates used in the enzyme assay. The mathematical models describing the single substrate as well as the mutual influence of phenol and p-cresol mixture were developed on the bases of Haldane kinetics. The interaction coefficients (Iph/cr, Icr/ph) indicated the degree to which phenol affects the biodegradation of cresol and vice versa were estimated.

Bioprospection of underexplored ecosystems have been proven as useful habitats for exploiting numerous bioactive metabolites from novel actinomycetes (Shah et al., 2017). Actinomycetes are known for the production of extracellular enzymes with applications in agriculture and industries (Mukhtar et al.,

Recent metagenomic studies have revealed that microbial diversity in the atmosphere rivals that of surface environments. This indicates that the atmosphere may be worth bioprospecting in for novel microorganisms, especially those selected for by harsh atmospheric conditions. This is interesting in light of the antibiotic resistance crisis and renewed interests in bioprospecting for members of the Actinomycetales, which harbor novel secondary metabolite-producing pathways and produce spores that make them well suited for atmospheric travel. The latter leads to the hypothesis that the atmosphere may be a promising environment in which to search for novel Actinomycetales. Although ubiquitous in soils, where bioprospecting efforts for Actinomycetales have been and are largely still focused, we present novel data indicating that culturable members of this taxonomic order are 3-5.6 times more abundant in air samples collected at 1.5, 4.5, 7.5, and 18 m above the ground, than in the underlying soil. These results support the hypothesis that mining the vast and readily accessible lower atmosphere for novel Actinomycetales in the search for undescribed secondary metabolites could prove fruitful.

Nowadays gelatinase have received considerable attention as targets for drug development because of their potential role in connective tissue degradation associated with tumor metastasis. The potential uses of Gelatinase and their high demand, necessitates the need for the discovery of new strains of bacteria that produce enzymes with novel properties and with low cost industrial medium. The present study was conducted to isolate and identify gelatinase producing microorganisms as well as optimize the medium and cultural conditions for maximum enzyme production. Isolates obtained from fish processing wastes were examined for gelatinase production.The medium parameters were optimized for maximum production of gelatinase enzyme by the efficient isolate.Optimizing its abiotic factors, maximum enzyme activity was achieved at 48h incubation period, pH 7, 40°C temperature, lactose as carbon source and a gelatin concentration of 0.2%. The identity of the strain was confirmed by PCR reaction with universal primers (27F and 1492 R), sequencing and then by the BLAST analysis. The results showed that the isolated bacteria had highest homology (100%) with Bacillus thuringiensis strain O3.The enzyme produced under all the optimum was then purified by ammonium sulphate precipitation (40%) and dialysis. The final purification after dialysis yielded specific activity of 45.85 IU/ml. The purity of enzyme preparation was assessed by SDS PAGE Gel electrophoresis and was found to be a single band with a molecular weight of approximately 66 kDa.The study could isolate and identify a potent gelatinase producing Bacillus strain that can be used for the development of gelatinase enzyme production.

The present study deals with the production of alpha amylase by Penicillium sp. employing solid state fermentation. Seven different media were evaluated. Out of all media tested wheat bran with distilled water gave maximum production of alpha amylase. Different parameters such as substrate concentration, moisture content, temperature, incubation time, size of inoculum and pH were optimized for the enzyme production. The maximum enzyme production was obtained by using 7.5 g of wheat bran, 100% of moisture content, incubation period 72 h, temperature of 30°C, inoculum size of 13.3% and pH 6. Addition of 1.5% starch as carbon source and 0.2% NH 4 NO 3 and 2% yeast extract as inorganic and organic nitrogen sources respectively gave maximum production of alpha amylase. In addition to this effect of metal ions was also investigated CaCl 2 act as stimulator for enzyme production by Penicillium sp.

All Idiomarina species are isolated from saline environments; microorganisms in such extreme habitats develop metabolic adaptations and can produce compounds such as proteases with an industrial potential. ARDRA and 16S rRNA gene sequencing are established methods for performing phylogenetic analysis and taxonomic identification. However, 16S-23S ITS is more variable than the 16S rRNA gene within a genus, and is therefore, used as a marker to achieve a more precise identification. In this study, ten protease producing Idiomarina strains isolated from the Peruvian salterns were characterized using biochemical and molecular methods to determine their bacterial diversity and industrial potential. In addition, comparison between the length and nucleotide sequences of a 16S-23S ITS region allowed us to assess the inter and intraspecies variability. Based on the 16S rRNA gene, two species of Idiomarina were identified (I. zobellii and I. fontislapidosi). However, biochemical tests revealed that there were differences between the strains of the same species. Moreover, it was found that the ITS contains two tRNA genes, tRNA Ile (GAT) and tRNA Ala (TGC) , which are separated by an ISR of a variable size between strains of I. zobellii. In one strain of I. zobellii (PM21), we found nonconserved nucleotides that were previously not reported in the tRNA Ala gene sequences of Idiomarina spp. Thus, based on the biochemical and molecular characteristics, we can conclude that protease producing Idiomarina strains have industrial potential; only two I. zobellii strains (PM48 and PM72) exhibited the same properties. The differences between the other strains could be explained by the presence of subspecies.

This study aims to optimize factors that affect power generation in microbial fuel cell (MFC) using a statistical experimental design. A MFC reactor with two compartments has been constructed from cheap materials and used for electricity generation. The performance of MFCs can be influenced by several factors, hence; the Plackett-Burman design was employed to determine the most significant factors that affect power generation. The factors that affected the electricity generation significantly were magnesium sulfate, yeast extract, and sodium acetate. A near optimum medium formulation was obtained using this method with increased power density by 71.57 %. In this respect, the three levels Box-Behnken design was applied. A polynomial model was created to correlate the relationship between the three significant variables and power density. Under the optimized condition, the power density was 390 mW/m 2. These results indicate that the optimized conditions accelerated the power generation and the maximum power point (MPP) was about 91.2 % higher than that recorded with the basal condition. Microbial fuel cells (MFCs) are new types of bioreactors that use bacteria to generate electricity from organic and nonorganic compounds and are considered one of the prospective alternative technologies.

Dependence on foreign oil remains a serious issue for the U.S. economy. Additionally, automobile emissions related to petroleum-based, fossil fuel has been cited as one source of environmental problems, such as globalwarming and reduced air quality. Using agricultural and forest biomass as a source for the biofuel ethanol industry, provides a partial solution by displacing some fossil fuels. However, the use of high cost enzymes as an input is a significant limitation for ethanol production. Economic analyses of cellulase enzyme production costs using solid state cultivation (SSC) are performed and compared to the traditional submerged fermentation (SmF) method. Results from this study indicate that the unit costs for the cellulase enzyme production are $15.67 per kilogram ($/kg)and $40.36/kg, for the SSC and SmF methods, respectively, while the market price for the cellulase enzyme is $36.00/kg. Profitability analysis and sensitivity analysis also provide positive results. Since th...

As many as 71 marine bacterial DNA extracts were PCR screened for L‐asparaginase (ansA), a key gene in anti‐cancer molecular‐searches. Over 62% (44) of them were positive for ansA gene. The positive cultures were from genera Bacillus and Staphylococcus. The ansA gene cloned from isolate NIOS4 belonging to recently described Bacillus tequilensis is 1099 bp in length with a 990 bp ORF coding for 329 amino acids. BLASTx analysis revealed this sequence to be 98% similar to earlier reported ansA sequence from B. subtilis (Accession no. NP390239.1). By comparing its deduced amino acid sequence with other bacterial asparaginase sequences six substitutions at positions 305(Thr), 313(Lys), 314(Leu), 315(Asp), 318(Arg), and 320(Gln) are observed. Key residues like Thr(12), Thr(85), Asp(86), Lys(156), and Phe(165) taking part in active‐site formation and imparting catalytic properties are conserved. The phylogenetic tree based of the ansA amino acid sequences revealed close relatedness of the ...

Mangrove ecosystems are environments subject to substantial degradation by anthropogenic activities. Its location, in coastal area, interfacing the continents and the oceans makes it substantially important in the prospection for biotechnological applications. In this study, we assessed the diversity of culturable bacteria present over the seasons at two depths (0-10 and 30-40 cm) in a mangrove sediment and in a transect area from the land to the sea. In total, 238 bacteria were isolated, characterized by Amplified Ribosomal DNA Restriction Analysis (AR-DRA) and further identified, by Fatty Acid Methyl Esther (FAME-MIDI), into the orders of Vibrionales, Actinomycetales and Bacillales. Also the ability of the isolates in producing economically important enzymes (amylases, proteases, esterases and lipases) was evaluated and the order Vibrionales was the main enzymatic source.

Biocatalysts are biological substances that act as an accelerator of chemical reactions without causing influence on the living organism chemically. Biocatalysts based on renewable sources, biodegradable, tolerance to pH and temperature, and high selectivity to stereochemistry substrates and products. The utilization of biocatalyst is environmentally friendly and effective in production costs. This research aimed to determine the potential of actinomycetes as a biocatalyst in biodiesel production. This research method isolation and identification of actinomycetes isolates, conducting lipase activity test to determining lipase enzyme production of actinomycetes isolates. The data analysis in microbial identification was conducted by molecular identification by Gene bank through Basic Local Alignment Search Tool (BLAST). The lipase activity of actinomycetes analyzing by the value of lipase rate of test microbes. Based on chemical analysis of lipase activity, this research results show...

Both positive and negative interactions among bacteria take place in the environment. We hypothesize that the complexity of the substrate affects the way bacteria interact with greater cooperation in the presence of recalcitrant substrate. We isolated lignocellulolytic bacteria from salt marsh detritus and compared the growth, metabolic activity and enzyme production of pure cultures to those of three-species mixed cultures in lignocellulose and glucose media. Synergistic growth was common in lignocellulose medium containing carboxyl methyl cellulose, xylan and lignin but absent in glucose medium. Bacterial synergism promoted metabolic activity in synergistic mixed cultures but not the maximal growth rate (μ). Bacterial synergism also promoted the production of β-1,4-glucosidase but not the production of cellobiohydrolase or β-1,4xylosidase. Our results suggest that the chemical complexity of the substrate affects the way bacteria interact. While a complex substrate such as lignocellulose promotes positive interactions and synergistic growth, a labile substrate such as glucose promotes negative interactions and competition. Synergistic interactions among indigenous bacteria are suggested to be important in promoting lignocellulose degradation in the environment.

The mixture of phenol and 4-methylphenol (p-cresol) effect on the growth behavior and degradation capacity of Aspergillus awamori strain was investigated. The phenol hydroxylase activity modulation depending on substrates included in the media was established. The enzyme hydroxylation of benzene-1,4-diol (hydroquinone) or 4-methylbenzene-1,2-diol (p-methylcatechol) goes better compared with phenol or p-cresol substrates used in the enzyme assay. The mathematical models describing the single substrate as well as the mutual influence of phenol and p-cresol mixture were developed on the bases of Haldane kinetics. The interaction coefficients (Iph/cr, Icr/ph) indicated the degree to which phenol affects the biodegradation of cresol and vice versa were estimated.

In order to obtain different actinomycetes isolates from the East Black Sea Region plateau soil, an isolation study was carried out by using sucrose gradient method and different growth media. A molecular taxonomic method, 16S rDNA gene sequencing, was applied to obtain different actinomycetes isolates from East Black Sea region plateau soil. Phylogenetic dendrograms based on 16S rDNA nucleotide sequences were constructed by using neighbour-joining algorithm. In the light of phylogenetic analyses, it was determined that out of 15 organisms, two belong to Actinomadura, three Kribbella, three Nocardia, six Micromonospora and an organism of Microbacterium. Soil isolates obtained from this study contribute to taxonomy of actinomycetes and also many species will emerge.

Las xilanasas son enzimas que hidrolizan el xilano, produciendo azúcares fermentables tales como xilosa y xilobiosa con aplicaciones industriales muy diversas entre las que destaca el biobleaching y la producción de bioetanol de segunda generación. El objetivo principal del presente estudio se enfoca en la optimización de la producción de xilanasa bacteriana por parte de la bacteria Bacillus sp. K1. La misma ha sido previamente aislada de muestras de madera en putrefacción recolectadas en las premisas de la Universidad de Lakehead para su Departamento de Biología. Las variables investigadas para la optimización fueron físicas tales como pH, temperatura y volumen de inóculo; y de composición del medio de fermentación tales como fuente de carbono, fuente de nitrógeno orgánico e inorgánico y relación porcentual entre dichos componentes. Adicionalmente, se determinó el efecto de iones metálicos (sodio, potasio, magnesio, calcio, ferroso, niqueloso, cúprico, cobaltoso y manganoso) y surf...

A fungal strain belonging to the genus Penicillium was isolated from soil sample and has been diagnosed as Penicillium sp. according to its morphological characteristics of the colonies on solid media and also microscopical examination of the fungal parts. Antibiotics, protease activity and pH values were determined after cultivation of the fungus using submerged fermentation (SF) and solid state fermentation (SSF). The two different patterns of fermentation processes seem to influence the physiological behavior of the fungus differently. Experiments were made using nutrient broth medium (N.B) for SF and wheat bran in SSF. The pH values were adjacent to 5.5. Wheat bran was enriched with fish scales and egg shale in a ratio of (1:2:0.005 w/w) and the mixture was moistened by adding (30 ml) whey solution. After 7 days of incubation, the pH value of SF was increased to 8.0 at 30ºC. The SF was appeared efficient for antibiotics production. Using well diffusion technique the extracted antibiotics solution was active against some pathogenic bacteria such as Staphylococcus aureus, E. coli, Proteus sp., Salmonella sp., Pseudomonas aeruginosa and Streptococcus sp. In SSF relative proteases concentrations were found to be highly reactive than SF. This was proved by the appearance of the zone (20 mm and 32 mm) due to the hydrolysis of milk and blood proteins respectively using pH 5.5 at 30ºC for 24 hrs. The activity of proteases was (10.4 U/ml).

Triticale (Triticosecale, Wittmak) an important industrial crop, with high grain yield, containing high amounts of starch, proteins and also major and minor mineral elements, is not sufficiently utilized yet. The simultaneous production of ?-amylase and glucoamylase isoforms by Aspergillus niger on triticale grains, without any nutritive supplements, has been developed, optimized and scaled up 10 fold for the first time. The specific combination of examined effects led to the production of a novel glucoamylase isoform. Reduction of particle size, increase of oxygen availability and substrate height lead to increase of amylases production of 30%. Reduction of relative humidity from 65% to 30% increased glucoamylase production 2 times and ?-amylase production for 30%. Peak production of ?-amylase (158 U/g) and glucoamylase (170 U/g) were obtained in erlenmayers and in scaled-up trays. The obtained A. niger amylase cocktail was more efficient in raw starch hydrolysis from wheat flour, ...

For the purpose of finding a suitable available inducer in combination with starvation, carbohydrate mixtures from triticale was used as inducers and compared with well-known amylase inducers in fungi. Carbohydrate mixtures from triticale induced production of amylase cocktail (?-amylase and glucoamylase) in Aspergillus niger, unlike induction with well-known inducers which induce only glucoamylase, showed by zymogram and TLC analysis of carbohydrates mixtures before and after fermentations. Glucoamylase production by A. niger was highest in the presence of extract obtained after autohydrolysis of starch from triticale (95.88 U/mL). Carbohydrate mixtures from triticale induced production of ?-amylase in A. oryzae. More ?-amylase isoforms were detected upon using complex carbohydrate mixture, compared to induction with maltose or starch. The 48 h induction was the most efficient by using triticale extract (101.35 U/mL). Carbohydrates from triticale extracts can be used as very good c...

Bacterial cellulose (BC) is a highly crystalline and mechanically stable nanopolymer, which has excellent potential as a material in many novel applications, especially if it can be produced in large amounts from an inexpensive feedstock. Traditionally, BC is produced from expensive culture media, containing glucose as carbon source and other nutrient sources resulting in very high production costs, which limits the use of this material to very high value added applications. The use of cheap carbon and nutrient sources is an interesting strategy to overcome this limitation and therefore to increase the competitiveness of this unique material. Waste papers as well as coconut water, rich source of carbon is used for economical production of BC. In this study, the possibility to combine production of BC and hydrolytic enzymes from waste was investigated. Bacterial cellulose and enzymes were produced through sequential saccharification and fermentations with the filamentous mixed fungal cultures and the bacterium Gluconacetobacter xylinus. The BC yield in paper hydrolysate based medium reached 7.2 g/l and in coconut water medium reached 8.3 g/l after 4 days of cultivation but in medium containing mixture of coconut water and paper hydrolysate the BC yield was increased to 10 g/l. The cellulase enzyme was produced from bacterial cellulose production medium and was found to be 1.02 U/ml. It was shown that waste paper as well as coconut water is a suitable raw material for production of bacterial cellulose and enzymes through sequential fermentation. The concept studied offers efficient utilization of the various waste materials and affords a possibility to combine production of two high valueadded materials.

A 20 kDa antifungal serine protease from Streptomyces sp. A6 was purified to 34.56 folds by gel permeation chromatography. The enzyme exhibited highest activity at neutral to near alkaline pH 7-9 and 55 °C. Neutral surfactant triton X-100 enhanced the activity by 4.12 fold. The protease activity also increased (109.9-119%) with increasing concentration of urea (2-8 mole/l). The enzyme was identified as serine protease with 67% similarity to SFase 2 of Streptomyces fradiae by MALDI-LC-MS/MS analysis. Determination of kinetic constants k m , V max , k cat and k cat /k m suggested higher affinity of enzyme for N-Suc-Ala-Ala-Val-Ala-pNA (synthetic substrate for chymotrypsin activity). The enzyme was highly stable at temperature prevailing under field conditions (40 °C) as apparent from K d and t 1/2 values, 0.0065 and 106.75 min, respectively and high ΔG* and negative ΔS* values, 87.17 KJ/mole and-126.95 J/mole, respectively. Thermal stability and increased activity of protease in presence of commonly used chemical fertilizer, urea, suggested its feasibility for agricultural applications. The present study is the first report on thermodynamic and kinetic properties of an antifungal protease from Streptomyces sp. A6. The study reflects potential of this enzyme for biocontrol of fungal plant pathogens.

A large number of actinomycetes (hundreds, thousands, and millions of CFU/g of soil) were iso lated from saline soils in the territory of the landscapes of the Ber hillocks. A significantly smaller quantity of actinomycetes was isolated from the soda saline soils and sor saline soils that formed at the bottom of the ephemeral salty lakes in Buryatiya and in the estuary of the Syr Darya River. Actinomycetes were represented in the studied soils by the Streptomyces, Micromonospora, Actinomadura, and Nocardiopsis genera. Among streptomycetes, the species of the Albus section and the Albus series predominated. The activity of the con sumption of substrates by cultures of actinomycetes was largely influenced by conditions of preincubation. Obviously, the influence was related to the alteration in the metabolism of actinomycetes as one of the mech anisms of adaptation to the increased osmolarity of environment. The alcalotolerance, thermotolerance, and xerotolerance of halotolerant actinomycetes of the saline soils of arid territories were experimentally revealed.

The glucose oxidase enzyme (GOx), also known as notatin (EC number 1.1.3.4), functions as an oxidoreductase that facilitates the oxidation of glucose to hydrogen peroxide and D-glucono-δ-lactone. This lactone then spontaneously converts to gluconic acid while concurrently generating hydrogen peroxide. The widespread applications of glucose oxidase have garnered significant attention across diverse industries such as chemical, pharmaceutical, food and beverage, clinical chemistry, biotechnology, and others. This enzyme is naturally produced by certain species of fungi and insects and exhibits antibacterial properties in the presence of oxygen and glucose. As the demand for glucose oxidase continues to rise and its production remains limited and costly, the reuse of enzymes becomes crucial. Moreover, when the enzymes are immobilized on suitable surfaces, their activity tends to increase. Immobilization refers to the process of attaching enzymes to surfaces, and various methods exist for achieving this immobilization. The choice of a specific immobilization method depends on factors such as the intended application, cost, activity, reproducibility, and half-life. This review provides a comprehensive overview of the principles and methods of enzyme immobilization in general and specifically focuses on a chronological survey of immobilized glucose oxidase. It further discusses the objectives and applications associated with this field.

Evidence from the available literature identified the potential use of filamentous fungi for the treatment of the cassava starch processing wastewater. Experimentation was also done on eight strains of filamentous fungi, which were known as producers of amylase and glucoamylase for digesting raw starch. Aspergillus oryzae strains showed high efficiency in the removal of COD and TOC, starch hydrolysis and production of fungal biomass in the medium of the synthetic cassava starch processing (SCSP) wastewater containing pure cassava starch. With A. oryzae KCC FOOI0, treatment efficiency obtained 72% (TOC), 81 % (COD), 94 % (starch) and 2.4 gil of biomass after 14-day incubation. The presence of cyanide caused inhibition for the growth of filamentous fungi. The toleration of A. oryzae IFO 30113 to cyanide was associated with its ability to remove cyanide. Ammonia was indicated as the end product of biodegradation of cyanide. Volatilization and microbial metabolism could be considered as...

Bacterial cellulose (BC) is a highly crystalline and mechanically stable nanopolymer, which has excellent potential as a material in many novel applications, especially if it can be produced in large amounts from an inexpensive feedstock. Traditionally, BC is produced from expensive culture media, containing glucose as carbon source and other nutrient sources resulting in very high production costs, which limits the use of this material to very high value added applications. The use of cheap carbon and nutrient sources is an interesting strategy to overcome this limitation and therefore to increase the competitiveness of this unique material. Waste papers as well as coconut water, rich source of carbon is used for economical production of BC. In this study, the possibility to combine production of BC and hydrolytic enzymes from waste was investigated. Bacterial cellulose and enzymes were produced through sequential saccharification and fermentations with the filamentous mixed fungal cultures and the bacterium Gluconacetobacter xylinus. The BC yield in paper hydrolysate based medium reached 7.2 g/l and in coconut water medium reached 8.3 g/l after 4 days of cultivation but in medium containing mixture of coconut water and paper hydrolysate the BC yield was increased to 10 g/l. The cellulase enzyme was produced from bacterial cellulose production medium and was found to be 1.02 U/ml. It was shown that waste paper as well as coconut water is a suitable raw material for production of bacterial cellulose and enzymes through sequential fermentation. The concept studied offers efficient utilization of the various waste materials and affords a possibility to combine production of two high valueadded materials.

The present study evaluates possibility to produce biocellulose nanofibers (BC) in simultaneous saccharification and fermentation (SSF). Pre-treated wheat straws (WS) were further incubated in the fermentation broth in the presence of Gluconacetobacter xylinus bacterium in the presence and absence of hydrolysis enzymes. WS were not filtered as common during separate hydrolysis and fermentation (SHF). Generally, results in the present study demonstrate that BC production in SSF is rather challenging, especially in the presence of hydrolysis enzymes. Total sugars produced during SSF were higher than SHF, and were generally identical under different pre-treatment and hydrolysis conditions (~54 g/L). This represents maximum amounts at complete hydrolysis of biomass due to the longer incubation time compared to SHF. Maximum BC production of 10.8 g/L was achieved when WS was chemically pretreated with 1% (by volume) dilute acid for 30 minutes at 121°C. Sample pre-treated with 2% acid at s...

The white rot fungus P. eryngii was grown on chemically and hot water treated agrowaste Brassica haulms. The fungus degraded lignin, cellulose, hemicellulose and carbon content of both chemically as well as hot water treated waste and produced in turn the edible and nutritious fruiting body. The progressive breakdown of lignin, cellulose and hemicellulose was correlated with apparent increase in the activities of lignolytic, cellulolytic and hemicellulolytic enzymes. Lignin degraded at faster rate during the vegetative phase and at slower rate during reproductive phase whereas, cellulose and hemicellulose depleted slowly during vegetative phase and rapidly during reproductive phase. The carbon content of the agrowaste decreased while, the nitrogen content increased and the C/N ratio came closer during degradation of the waste. Hot water treated substrate supported better production of enzymatic activity and degraded more efficiently than chemically sterilized substrate. The total yi...