Increase In Enzyme Production Research Articles

Leucoanthocyanidin Reductase 3 (PaLAR3) Locus in Norway Spruce (Picea abies) and Its Link to Resistance Against Heterobasidion parviporum

ABSTRACTHeterobasidion parviporum is a fungal pathogen that is drastically damaging Norway spruce (Picea abies) in Europe. The infections will result in root and stem rot, causing significant economic losses for forest owners. Previous studies have shown that the PaLAR3 gene, which encodes the leucoanthocyanidin reductase enzyme, can increase resistance to H. parviporum in Norway spruce. The presence of the B allele at the PaLAR3 locus has been associated with higher (+)‐catechin concentrations and increased enzyme production in inoculation experiments, resulting in inhibited pathogen growth. The control of H. parviporum involves a multifaceted approach, including silvicultural and sustainable forestry practices, genetic resistance and chemical/biological control. In this study, we determined the PaLAR3 genotypes in a representative sample of Norway spruce breeding materials from southern Finland and examined their effect on necrosis caused by H. parviporum in spruce stems. The results showed that the homozygous PaLAR3BB genotype was present in only 9% of the trees. However, the necrotic area interacted with homozygous PaLAR3BB under low‐water treatment. These findings support the idea that the PaLAR3 locus may be a valuable marker for identifying P. abies resistance to different strains of Heterobasidion parviporum.

Combinational manipulation of transcription factors, CreA and ClbR, is a viable strategy to improve cellulolytic enzyme production in Aspergillus aculeatus

The production of cellulolytic enzymes in response to inducible carbon sources is mainly regulated at the transcriptional level in filamentous fungi. We have identified a cellobiose-response regulator (ClbR) controlling the expression of cellulolytic enzyme-encoding genes in Aspergillus aculeatus. However, the engineering potential of combining the deletion of transcriptional repressors with the overexpression of transcriptional activators to enhance enzyme production has not been analyzed. Here, we investigated the effect of the deletion of the transcriptional repressor creA and the overexpression of the transcriptional activator clbR in enzyme production in A. aculeatus. Here, we verified that a combination of creA deletion and clbR overexpression (Δc&OE) improved cellulase, β-1,4-xylanase, and β-glucosidase production. Cellulase and β-1,4-xylanase production increased 3.4- and 8.0-fold in Δc&OE compared with the host strain (MR12) at 96-h incubation, respectively. β-Glucosidase production in ΔcreA and Δc&OE increased approximately 5.0-fold compared with that in MR12 at 240 h-incubation. Transcriptional analysis revealed that the increase in enzyme production was due to increased expression of cellobiohydrolase, endo-β-1,4-glucanase, β-1,4-xylanase, and β-glucosidase 1 (bgl1). Interestingly, bgl1 expression in ΔcreA increased in a dose-dependent manner in response to glucose. Thus, combinational manipulation of transcription factors improved cellulase, xylanase, and β-glucosidase production in A. aculeatus.

Creating a novel genetic diversity of Trichoderma afroharzianum by γ-radiation for xylanase-cellulase production

Creating novel sources of a microbial strain using induced mutation can increase enzyme production for industrial use. According to this, we have developed a mutant strain of Trichoderma afroharzianum by Co60 gamma irradiation. Trichoderma mutants were isolated from an optimum dose of 250 Gy. The qualitative and quantitative screening were used for evaluating their enzyme production and the DNA barcoding method was used to identify the best Trichoderma mutant isolates. The highest cellulase (exo-glucanase, endoglucanase, β-glucosidase, and total cellulase) and xylanase activities were observed in superior mutant isolates of Trichoderma afroharzianum NAS107-M44 and Trichoderma afroharzianum NAS107-M82, which is approximately 1.6–2.5 times higher than its parent strain, respectively. The electrophoretic pattern of proteins showed that the exo-glucanase I, endo-glucanase III, and the xylanase I enzymes hydrolyzed the corn bran, synergistically. Overall, gamma irradiation-induced mutation could be an expedient technique to access such superior mutants for the bioconversion of corn bran wastes.

In Vitro Microevolution and Co-Selection Assessment of Amoxicillin and Cefotaxime Impact on Escherichia coli Resistance Development.

The global spread of antimicrobial resistance has become a prominent issue in both veterinary and public health in the 21st century. The extensive use of amoxicillin, a beta-lactam antibiotic, and consequent resistance development are particularly alarming in food-producing animals, with a focus on the swine and poultry sectors. Another beta-lactam, cefotaxime, is widely utilized in human medicine, where the escalating resistance to third- and fourth-generation cephalosporins is a major concern. The aim of this study was to simulate the development of phenotypic and genotypic resistance to beta-lactam antibiotics, focusing on amoxicillin and cefotaxime. The investigation of the minimal inhibitory concentrations (MIC) of antibiotics was performed at 1×, 10×, 100×, and 1000× concentrations using the modified microbial evolution and growth arena (MEGA-plate) method. Our results indicate that amoxicillin significantly increased the MIC values of several tested antibiotics, except for oxytetracycline and florfenicol. In the case of cefotaxime, this increase was observed in all classes. A total of 44 antimicrobial resistance genes were identified in all samples. Chromosomal point mutations, particularly concerning cefotaxime, revealed numerous complex mutations, deletions, insertions, and single nucleotide polymorphisms (SNPs) that were not experienced in the case of amoxicillin. The findings suggest that, regarding amoxicillin, the point mutation of the acrB gene could explain the observed MIC value increases due to the heightened activity of the acrAB-tolC efflux pump system. However, under the influence of cefotaxime, more intricate processes occurred, including complex amino acid substitutions in the ampC gene promoter region, increased enzyme production induced by amino acid substitutions and SNPs, as well as mutations in the acrR and robA repressor genes that heightened the activity of the acrAB-tolC efflux pump system. These changes may contribute to the significant MIC increases observed for all tested antibiotics. The results underscore the importance of understanding cross-resistance development between individual drugs when choosing clinical alternative drugs. The point mutations in the mdtB and emrR genes may also contribute to the increased activity of the mdtABC-tolC and emrAB-tolC pump systems against all tested antibiotics. The exceptionally high mutation rate induced by cephalosporins justifies further investigations to clarify the exact mechanism behind.

Thermoalkalophilic polygalacturonase from a novel Glutamicibacter sp.: Bioprospecting, strain improvement, statistical optimization and applications

The current research discusses a multidimensional bioprocess development, that includes bioprospecting, strain improvement, media optimisation, and applications of the extracted enzyme. A potent alkalophilic polygalacturonase (PG) producing bacterial strain was isolated and identified as a novel Glutamicibacter sp. Furthermore, strain improvement by UV and chemical mutagenesis not only improved the enzyme (PGmut) production but also enhanced its temperature optima from 37 °C to 50 °C. The use of solid substrate fermentation, followed bystatistical optimisation through PB and RSM, substantially increasedPGmut production. A 10-fold increase in enzyme production (632 U/gm) was observed when sugarcane bagasse with a pH of 10.5, 66.8 % moisture, and an inoculum size of 10.15 % was used. The model’s accuracy was supported by p-value (p < 0.0001), and an R2 of 0.9940. A pilot-scale experiment, demonstrated ≈ 62,229 U/100 gm PG activity. Additionally, the enzyme’s efficacy in demucilization of coffee beans, and bioscouring of jute fibre indicated that it is a valuable biocatalyst.

Scale-up studies for production of α-amylase by A. Tamarii MTCC5152 and development of enzyme product

α-amylases are one of the most vital commercially produced starch hydrolyzing enzymes. Increasing enzyme production is becoming one of the most important goals in biotechnological processes. In this study, the production of α-amylase by A.tamarii MTCC5152 in flask level studies is experimentally validated by scale-up studies in Koji room under controlled conditions using wheat bran, a low cost agro-industrial waste as substrate. The production of amylase was increased by optimising various culture conditions such as inoculum size, incubation period and moisture content. It was found that A.tamarii MTCC5152 produced highest titre of α-amylase activity (668.01 u/g), after 4 days of incubation period at 5% inoculum concentration and 80% moisture content. The results of the comparative analysis of protease and amylase production demonstrated 15% increase in α- amylase and 20% increase in protease production compared to flask level studies. Studies conducted for the development of enzyme products as liquid enzyme concentrate and dry enzyme powder showed that addition of hydrophilic agents such as poly ethylene glycol (PEG- 6000) was found to be an effective way to increase the activity and enzyme yield in the preparation of a dry enzyme powder using A.tamarii MTCC5152. The ammonium sulfate precipitated enzyme at saturation, lyophilized by freeze drying gave the best recovery (96%) of the enzyme activity.

Bentonite Impact on Soil Properties and Biological Activity in the Face of Drought : A Review

The clay mineral bentonite has received a lot of attention for its potential to reduce the harmful effects of drought on soil health and biological activity. Reduced enzymatic activity and microbial diversity are two negative outcomes of the drought’s impact on soil quality. Bentonite, however, has been shown to aid in the restoration of biological functions in drought-affected soils. Bentonite’s function as a soil moisture retainer is an important consideration. Bentonite acts as a reservoir, trapping water within its layers due to its high water-holding capacity and ability to form a gel-like structure. This capacity for water retention can reduce the negative impact of drought on soil organisms, making conditions better for enzyme activity and microbial life. Bentonite has also been shown to increase soil enzyme production and activity. Soil fertility and healthy ecosystems rely on the work of enzymes in nutrient cycling and organic matter decomposition. Reduced enzymatic activity is a common consequence of drought stress, which in turn reduces nutrient availability and worsens soil health. The application of bentonite, however, has been shown to increase enzyme production and activity even in drought conditions. This has the potential to improve soil health and productivity by preserving vital soil processes and encouraging nutrient cycling. In addition, bentonite can increase soil microorganism activity in dry areas. Soil microorganisms play a crucial role in the cycling of nutrients, the control of disease, and the health of the soil ecosystem as a whole. Reduced microbial diversity and activity have been linked to drought stress. However, research shows that applying bentonite to drought-affected soils increases both microbial growth and diversity. Increased nutrient availability, enhanced soil structure, and enhanced water infiltration and retention are all possible results of microbial stimulation. In conclusion, areas that have experienced drought can benefit from bentonite’s ability to improve soil enzymes and biological activity. Bentonite helps restore and maintain soil health even when water supplies are low. It does this by increasing water retention, encouraging enzyme production, and boosting microbial activity. To fully realize bentonite’s potential for fostering sustainable agriculture and sustaining healthy ecosystems, more study is required to determine the optimum application rates and methods.

Bioprospecting of a Thermostable L-Methioninase from Alcaligenes aquatilis BJ-1 in Agro-Industrial Waste

L-methioninase is an enzyme that has recently gained significant interest in the scientific community because of its potential as a targeted therapy for cancer. This study aims to isolate and identify extremophilic bacteria that could produce L-methioninase and to access the enzymatic potential of isolated bacteria under stress conditions, specifically in agro-industrial waste. In this study, a rare marine bacterium, Alcaligenes aquatilis BJ-1, exhibited the highest specific activity of 4.61 U/mg at an optimum pH of 8.3. The L-methioninase was purified 4.3-fold and 7.15-fold by acetone precipitation and Sephadex G-100 gel filtration chromatography, which revealed a molecular weight of 46 kDa. In addition, agriculture waste materials such as cottonseed oil cake had the highest L-methioninase production. Moreover, A. aquatilis BJ-1 can tolerate and produce enzymes in the presence of 10% NaCl, 6% KCl, and 4% MgSO4. Similarly, substrates such as L-asparagine, L-glutamine, L-alanine, and L-tyrosine were found suitable to increase enzyme production. The strain produced L-methioninase in the presence of various heavy metals. Maximum enzyme activity was found in Zn2+ at 0.1% (2.52 U/mL), Li2+ at 0.03% (2.90 U/mL), and Ni2+ at 0.01% (2.78 U/mL), as compared to the control (2.23 U/mL) without metal. Enzyme production was also observed at a high temperature (60 °C), with the produced enzymes possessing antioxidant properties. In addition, no hemolytic activity was observed. The results indicate that A. aquatilis BJ-1 is an appropriate bacterium for metal bioremediation procedures in unfavorable circumstances.

Current state of molecular and metabolic strategies for the improvement of L-asparaginase expression in heterologous systems.

Heterologous expression of L-asparaginase (L-ASNase) has become an important area of research due to its clinical and food industry applications. This review provides a comprehensive overview of the molecular and metabolic strategies that can be used to optimize the expression of L-ASNase in heterologous systems. This article describes various approaches that have been employed to increase enzyme production, including the use of molecular tools, strain engineering, and in silico optimization. The review article highlights the critical role that rational design plays in achieving successful heterologous expression and underscores the challenges of large-scale production of L-ASNase, such as inadequate protein folding and the metabolic burden on host cells. Improved gene expression is shown to be achievable through the optimization of codon usage, synthetic promoters, transcription and translation regulation, and host strain improvement, among others. Additionally, this review provides a deep understanding of the enzymatic properties of L-ASNase and how this knowledge has been employed to enhance its properties and production. Finally, future trends in L-ASNase production, including the integration of CRISPR and machine learning tools are discussed. This work serves as a valuable resource for researchers looking to design effective heterologous expression systems for L-ASNase production as well as for enzymes production in general.

Assessment of cultivation parameters influencing pectinase production by Aspergillus niger LFP-1 in submerged fermentation

BackgroundPectinase is helpful in food and beverage industries, particularly in the preparation of fruit juice, the extraction of vegetable oil, and the fermentation of coffee. The current work aimed to screen Aspergillus niger LFP-1, a recently identified fungal strain, for its ability to produce pectinase and to ascertain the contribution of various physicochemical factors to pectinase production. ResultsThe primary and secondary pectinase activity screenings by Aspergillus niger LFP-1 were performed using pectin screening agar and shake flask system, respectively. The finding revealed that the locally isolated strain is able to secrete favourable pectinase production. Before improvement, the pectinase production was 0.88 ± 0.09 U/mL. However, the improved conditions such as 6 days of the cultivation period, agitation speed of 150 rpm, inoculum size of 1 × 106 spores/mL, 2.5% (w/v) citrus pectin, and 0.4% (w/v) ammonium nitrate could significantly increase pectinase production up to 7.41 ± 0.24 U/mL, representing an 88% increase. In this study, supplementing 2.5% (w/v) citrus pectin to the culture medium as a carbon source increased enzyme production by up to 3.07 ± 0.17 U/mL. Meanwhile, 0.4% (w/v) ammonium nitrate was used as a nitrogen source yielding the highest enzyme activity with a value of 6.86 ± 0.07 U/mL. ConclusionThus, the locally isolated fungal strain, A. niger LFP-1 has outstanding pectinase-producing capability and can be utilized for the commercial production of pectinase. The improved cultural conditions significantly increase pectinase production and shorten the incubation period from 8 days (before improvement) to 6 days (after improvement).

Genetic Fidelity Studies using RAPD Marker on local fungal isolate of Jabalpur Region for over-expression of Glucoamylase

The present study was conducted to study genomic profile of wild and mutant isolates of local soil fungal isolate Aspergillus flavipes strain for over-production of glucoamylase using RAPD markers. The study involves application of mutational strategies using chemical and physical mutagens on the potent fungal isolate and their screening studies thereafter. The alteration in genetic makeup of the mutant fungal isolate (SSP#16M) with the wild type (SSP#16) was detected and confirmed by RAPD-PCR analysis of genomic DNA. Although various genotypic methods have been used for comparison of Aspergillus species but no comparable data is available for Aspergillus flavipes strain for increased enzyme production. In the study, the mutant fungal isolate designated NA3 (exposed to nitrous acid) showed 3.5 fold improved glucoamylase activity than wild isolate and showed consistency in the production of the enzyme after repeated subcultures. The data for genetic variability was analysed using software Dendro UPGMA wherein the similarity matrix was computed with Jaccard coefficient. This assessment study of mutant isolate of Aspergillus flavipes strain provides potential applicability in starch processing industries.

Ocean acidification causes fundamental changes in the cellular metabolism of the Arctic copepod Calanus glacialis as detected by metabolomic analysis

Using a targeted metabolomic approach we investigated the effects of low seawater pH on energy metabolism in two late copepodite stages (CIV and CV) of the keystone Arctic copepod species Calanus glacialis. Exposure to decreasing seawater pH (from 8.0 to 7.0) caused increased ATP, ADP and NAD+ and decreased AMP concentrations in stage CIV, and increased ATP and phospho-L-arginine and decreased AMP concentrations in stage CV. Metabolic pathway enrichment analysis showed enrichment of the TCA cycle and a range of amino acid metabolic pathways in both stages. Concentrations of lactate, malate, fumarate and alpha-ketoglutarate (all involved in the TCA cycle) increased in stage CIV, whereas only alpha-ketoglutarate increased in stage CV. Based on the pattern of concentration changes in glucose, pyruvate, TCA cycle metabolites, and free amino acids, we hypothesise that ocean acidification will lead to a shift in energy production from carbohydrate metabolism in the glycolysis toward amino acid metabolism in the TCA cycle and oxidative phosphorylation in stage CIV. In stage CV, concentrations of most of the analysed free fatty acids increased, suggesting in particular that ocean acidification increases the metabolism of stored wax esters in this stage. Moreover, aminoacyl-tRNA biosynthesis was enriched in both stages indicating increased enzyme production to handle low pH stress.

Identification and Optimization Study of Lipase Producing Bacteria Isolated from Municipal Waste and Bio-deteriorated Waste

Bio-deteriorated waste is the leftover organic matter of unwanted raw food which if not handled properly or left for natural degradation can cause health issues. Microorganisms have the ability to biodegrade waste by secreting enzymes. The aim of the work was to isolate and identify lipase-producing bacteria from waste polluted (Bio-deteriorated waste and Municipal Solid Waste) dumping sites of Gandhinagar, Gujarat, India. Lipase-producing bacteria were isolated using tributyrin agar as a selective medium. Out of 7 bacterial isolates, 1 isolate (HAL-2) gave the highest lipolytic activity. HAL-2 was identified as Bacillus pumilus by 16S rRNA sequencing. The bacterial isolate gave maximum lipase activity (0.68 U/mL) at 37°C and pH 7.0 Culture medium parameters such as carbon source, nitrogen source, pH, and inoculum size were varied for the purpose of optimization. The maximum lipase production was observed at pH 7.0, 37°C temperature. Inoculum size had an effect of direct proportionality on lipase activity. Glycerol tributyrate was found to be the best substrate (0.68 U/mL). Sucrose and Tryptone in the medium increased enzyme production when compared with other carbon and nitrogen sources.

Statistical, physicochemical, and thermodynamic profiles of chitinase production from local agro-industrial wastes employing the honey isolate Aspergillus niger EM77

Enzyme synthesis from local wastes has a lot of potential because it eliminates the problem of waste accumulating in conjugation while also cutting the cost of these useful products. Plackett-Burman (PBD) and central composite designs (CCD) were used to optimize the manufacturing process utilizing the honey isolate Aspergillus niger EM77, resulting in a 60-fold increase in enzyme productivity using a group of wastes comprising wheat, rice straw, and sawdust. The enzyme had its optimum activity at 60 °C, pH 5, and had high thermo-stability at 60 °C, with Km and Vmax of 0.8 mg mL−1 and 2083.33 μmol mL−1 min−1 respectively. The activation (Ea) and deactivation (Ed) energies of chitinase were 2.78 and 174.46 kJ mol−1, respectively, with the thermodynamic constants ΔH° and ΔG° ensuring enzyme stability. As a result, Aspergillus niger EM77 chitinase has the efficiency to meet the global market demand for chitinase enzyme while also providing a significantly lower price than what is now available on the websites of specialist international companies. The production process is almost costless because it is based primarily on waste and contains traces of minerals.

Selection and Optimization of Medium Components for the Efficient Production of L-Asparaginase by Leucosporidium scottii L115—A Psychrotolerant Yeast

This study reports the production of L-asparaginase (ASNase), an enzyme mainly used for the treatment of acute lymphoblastic leukemia, by Leucosporidiumscottii L115, a psychrotolerant yeast isolated from the Antarctic ecosystem. Focus was given to select the most appropriate medium components able to maximize the enzyme production by this yeast, as a first step for the development of a new process to produce ASNase. By combining knowledge in bioprocesses, statistical analysis and modeling, the medium composition that most favored enzyme production was established, which consisted of using a mixture of sucrose (28.34 g L−1) and glycerol (15.61 g L−1) as carbon sources, supplemented with proline (6.15 g L−1) and the following salts (g L−1): KCl, 0.52; MgSO4·7H2O, 0.52; CuNO3·3H2O, 0.001; ZnSO4·7H2O, 0.001; and FeSO4·7H2O, 0.001. By using this medium, enzyme production of 2850 U L−1 (productivity of 23.75 U L−1 h−1) was obtained, which represented a 28-fold increase in enzyme production per gram of cells (178 U gdcw−1) when compared to the control (non-optimized medium), and a 50-fold increase when compared to a reference medium used for ASNase production.

Laccase production by Pleurotus ostreatus using cassava waste and its application in remediation of phenolic and polycyclic aromatic hydrocarbon-contaminated lignocellulosic biorefinery wastewater

The treatment of contaminants from lignocellulosic biorefinery effluent has recently been identified as a unique challenge. This study focuses on removing phenolic contaminants and polycyclic aromatic hydrocarbons (PAHs) from lignocellulosic biorefinery wastewater (BRW) applying a laccase-assisted approach. Cassava waste was used as a substrate to produce the maximum yield of laccase enzyme (3.9 U/g) from Pleurotus ostreatus. Among the different inducers supplemented, CuSO4 (0.5 mM) showed an eight-fold increase in enzyme production (30.8 U/g) after 240 h of incubation. The catalytic efficiency of laccase was observed as 128.7 ± 8.47 S−1mM−1 for syringaldazine oxidation at optimum pH 4.0 and 40 °C. Laccase activity was completely inhibited by lead (II) ion, mercury (II) ion, sodium dodecyl sulphate, sodium azide and 1,4 dithiothretiol and induced significantly by manganese (II) ion and rhamnolipid. After treating BRW with laccase, the concentrations of PAHs and phenolic contaminants of 1144 μg/L and 46160 μg/L were reduced to 96 μg/L and 16100 μg/L, respectively. The ability of laccase to effectively degrade PAHs in the presence of different phenolic compounds implies that phenolic contaminants may play a role in PAHs degradation. After 240 h, organic contaminants were removed from BRW in the following order: phenol >2,4-dinitrophenol > 2-methyl-4,6-dinitrophenol > 2,3,4,6-tetrachlorophenol > acenaphthene > fluorine > phenanthrene > fluoranthene > pyrene > anthracene > chrysene > naphthalene > benzo(a)anthracene > benzo(a)pyrene > benzo(b)fluoranthene > pentachlorophenol > indeno(1,2,3-cd)pyrene > benzo(j) fluoranthene > benzo[k]fluoranthène. The multiple contaminant remediation from the BRW by enzymatic method, clearly suggests that the laccase can be used as a bioremediation tool for the treatment of wastewater from various industries.

Effect of process parameters and surfactant additives on the obtained activity of recombinant tryptophan hydroxylase (TPH1) for enzymatic synthesis of 5-hydroxytryptophan (5-HTP)

5-hydroxytryptophan (5-HTP) is an intermediate molecule in the biosynthesis of serotonin, an important neurotransmitter, regulating a series of metabolic and psychological functions in humans. In this work, we studied the heterologous production of Human tryptophan hydroxylase (TPH1) in Escherichia coli, for the synthesis of 5-hydroxytryptophan (5-HTP) from Tryptophan (Trp). To quantify TPH1 activity, a simple fluorescence-based microtiter plate assay was established, based on the changes in fluorescence emission at 340 nm between substrate and product when excited at 310 nm, allowing quick and reliable quantification of released 5-HTP. To increase enzyme production, heterologous TPH1 production was studied in stirred tank bioreactor scale. The effect of rate of aeration (0.25, 0.50 and 0.75 vvm) and agitation (150, 250 and 500 rpm) was evaluated for biomass production, pH, volumetric oxygen transfer coefficient (kLa) and volumetric TPH1 activity. We determined that high agitation and low aeration allowed reaching the maximum measured enzyme activity. Under such conditions, we observed a 90% substrate conversion, obtaining 90 µM (~0.02 g/L) 5-HTP from a 100 µM Tryptophan substrate solution. Finally, we observed that the addition of Tween 20 (0.1%) in the culture broth under production conditions expanded the pH operation range of TPH1. Our results establish a base for a biocatalytic approach as a potential alternative process for the synthesis of 5-HTP using recombinant TPH1.

Preparation of Aspergillus niger 426 naringinases for debittering citrus juice utilization of agro-industrial residues.

Naringin, considered as the main bitter component of grapefruit, requires the use of enzymes to reduce the level of this substance during juice processing. For this reason, it has been the focus of many studies. In this study, to increase the production of naringinase by Aspergillus niger cultivated in solid-state fermentation (SSF), a three-component simplex-centric mixing design along with a response surface methodology (RSM) was applied to generate statistical models and analyze the dataset. First, grapefruit peel, rice bran, and wheat bran were used for substrate selection for naringinase production and, finally, selected the best of the three inducers or their mixtures to remove the bitterness of grapefruit juice. Cultivation with 2.3g of grapefruit peel, 2.5g of rice bran, and 5.2g of wheat bran and medium supplementation with a mixture of naringin, rutin, and hesperidin in the concentration of 2, 5, 4.5, and 3.0g/L, respectively, resulted in a maximum activity of 28 U/mL. The results indicate that the sequencing procedure, which allowed the definition of an optimal mixture of components, is a new way for microorganisms to have a high naringinase yield, in particular by SSF, since our data showed a 96% increase in the production of naringinase. This dataset can help other researchers apply a mixing design to increase enzyme production.

Lipase from new isolate Bacillus cereus ATA179: optimization of production conditions, partial purification, characterization and its potential in the detergent industry.

In this study, 341 Bacillus sp. strains were isolated from agricultural soils of Turkey. The potent extracellular lipase producer was selected. It was identified by 16S rRNA, named as Bacillus cereus ATA179. Optimal nutritional and physical parameters for lipase production were determined. Sucrose as the carbon source, (NH4)2HPO4 as the nitrogen source, CaCl2 as the metal ion were obtained. The best results of physical parameters were stated at 45°C, pH 7.0, shaking rate 50 rpm, inoculation amount 7% and inoculum age 24 h. ATA179 strain showed a 51% increase in enzyme production in the modified medium created by optimizing nutritional and physical conditions. Optimum pH value and temperature were found as 6.0 and 55 °C, respectively. CaCl2, Tween 20, Triton X-100 had an activating effect on enzyme activity. Vmax and Km kinetic values were found as 18.28 U/mL and 0.11 mM, respectively. The molecular weight was determined as 47 kDa. Lipase was found to be stable up to 75 days at -20 ºC. The potential of the enzyme in detergent industry was also investigated. It was not affected by detergent additives, but was found to be effective in removing oils from contaminated fabrics. This new lipase may have potential to be used in detergent industry.

Kinetics of growth and co-production of amylase and protease in novel marine actinomycete, Streptomyces lopnurensis KaM5.

Amylases and proteases are among the industrially most important enzymes for food processing, animal feed, brewing, starch processing, detergents, healthcare, leather processing, and biofuel production. In this study, we investigated the growth kinetics and statistically optimized the co-production of amylase and protease in a phylogenetically novel haloalkaliphilic actinomycete, Streptomyces lopnurensis KaM5 of seawater. The Plackett-Berman design using Minitab 14.0 software was employed to assess the impact of the nutritional factors, temperature, pH, and incubation time. Further, starch, yeast extract, NaCl concentrations, and incubation time were optimized by Box-Behnken design at their three levels. The Pareto charts, contour, surface plots, and individual factorial analysis expressed the variability and levels for the optimal enzyme production. ANOVA analysis admitted the statistical fitness and significance level among the variables. A two-fold increase in enzyme production was achieved by cost-effective co-production media. The study was further extended to growth kinetics associated with enzyme production. Specific growth rate (μ), maximal cell mass (Xmax), volumetric product formation (Pmax), rate of product formation (Qp), and generation time (g) were computed and analyzed. These parameters significantly improved when compared with the pre-optimized conditions, and the production economics of the enzyme was industrially viable. The initial studies on the characteristics of the enzymes suggested its ability to function under the combination of alkaline pH and high salt concentrations. The co-production of enzymes from extremophiles can be a potentially viable option for large-scale production and applications.