Activity Of Crude Enzyme Research Articles

Strategies for enhanced acetone-butanol-ethanol production by Clostridium beijerinckii Y10 from glucose and xylose and exploration of its physiological mechanisms

To enhance the production of butanol and its ratio to the total solvent, the impacts of exogenous amino acids supplementation on acetone-butanol-ethanol (ABE) fermentation were studied. With this effort, the highest butanol and total solvent production of 14.18 ± 0.24 g/L and 16.29 ± 0.31 g/L were achieved when 5.0 mg/L of phenylalanine was added at 18 h. Additionally, to explore the physiological reasons for the efficient synthesis of butanol under the optimal condition, the changes of key enzyme activities in the butanol synthesis pathway and the intracellular microenvironment were investigated. The findings show that phenylalanine addition could enhance crude enzyme activities of acetate kinase and butyrate kinase activities, strengthen NADH synthesis and expend more ATP to promote the growth of Clostridium beijerinckii Y10. This study suggests that the method of a tiny amount of amino acid addition was a simple and efficient approach to enhance bio-butanol production, thus providing a new strategy to improve the performance of other similar fermentation.

Production of thermostable recombinant cutinase using mixed food waste: A sustainable approach toward environmental remediation

Recently, there has been a significant surge in the utilization of single-use plastics. Cost-effective degradation of plastics is imperative to reduce the ecological burden. This study explores the overproduction and characterization of a PET (Polyethylene terephthalate) a degrading enzyme, cutinase, in the E. coli BL21 (DE3) expression system. Furthermore, this research delves into the utilization of mixed food waste (MFW) as a sustainable substrate for the production of Cutinase, replacing commercial glucose. The newly isolated strain, Aspergillus terreus, showed the highest glucoamylase activity of 76.10 ± 3.10 U/g. The enzymatic hydrolysis of mixed food waste resulted in 50 ± 0.4 g/L of glucose, used as feedstock for cutinase production. The crude enzyme activity was evaluated to be 11.64 U/mg, and a further 14.11-fold purification was achieved through IMAC. The purified recombinant enzyme showed its maximum activity at 70℃ and pH 7. The metal ion Na+ increased the enzyme activity by 26 % while Zn+2 significantly inhibited the same (47 % inhibition); other ions had little or no effect. Taguchi’s optimization of five variables showed that the concentration of food waste hydrolysate has the maximum impact on enzyme production. The treatment of commercially available PET bottles using the purified enzyme showed characterizable surface degradation effects. This is the first time food waste hydrolysate was used as a sole carbon source to produce recombinant cutinase. This approach can potentially pave the way for sustainable biorefinery and waste management.

Effect of medium supplementations and extraction conditions on cellulase production through solid state fermentation of oil palm empty fruit bunches

In this study, cellulase enzyme was produced through solid state fermentation (SSF), employing oil palm empty fruit bunches (EFB) as the primary substrate. Two key aspects were explored to enhance crude enzyme yield, which are medium supplementation effects during SSF and cellulase recovery. During medium supplementation, glucose and/or Tween 80 were added alongside EFB substrate and other nutrients. Enzyme yield was determined using a filter paper assay and expressed as enzyme activity. The initial addition of glucose during fermentation led to increased crude enzyme activity, as measured by the filter paper assay. The peak crude enzyme activity was observed with the addition of 3 mg of glucose, with higher amounts showing no further increase in activity. Conversely, the addition of Tween 80 did not yield any significant increase in crude enzyme activity across all concentrations tested. The extraction conditions were varied to study cellulase recovery, specifically by adjusting the solid‐to‐solvent ratio and the number of extraction stages. Higher enzyme activity was achieved with lower solid‐to‐liquid ratios, as the increased solvent volume facilitated greater enzyme extraction. However, increasing the number of extraction steps did not significantly affect the resulting cellulase activity. Overall, this research underscores the need for further process optimization for cellulase production via SSF, utilizing the widely available EFB in Indonesia.

Optimization of newly isolated Bacillus cereus α-amylase production using orange peels and crab shells and application in wastewater treatment.

A newly isolated amylolytic strain was identified as Bacillus cereus spH1 based on 16S and 16-23S gene sequencing (Accession numbers OP811441.1 and OP819558, respectively), optimization strategies, using one variable at time (OVAT) and Plackett-Burman design, were employed to improve the alpha-amylase (α-amylase) production. Condition inferred revealed that the optimal physical parameters for maximum enzyme production were 30°C, pH 7.5, and 12h of incubation, using tryptone, malt extract, orange (Citrus sinensis) peels, crab (Portunus segnis) shells, calcium, and sodium chloride (NaCl) as culture medium. The full factorial design (FFD) model was observed to possess a predicted R2 and adjusted R2 values of 0.9788 and 0.9862, respectively, and it can effectively predict the response variables (p = 0). Following such efforts, α-amylase activity was increased 141.6-folds, ranging from 0.06 to 8.5 U/mL. The ideal temperature and pH for the crude enzyme activity were 65°C and 7.5, respectively. The enzyme exhibited significant stability, with residual activity over 90% at 55°C. The maltose was the only product generated during the starch hydrolysis. Moreover, the Bacillus cereus spH1 strain and its α-amylase were used in the treatment of effluents from the pasta industry. Germination index percentages of 143% and 139% were achieved when using the treated effluent with α-amylase and the strain, respectively. This work proposes the valorization of agro-industrial residues to improve enzyme production and to develop a green and sustainable approach that holds great promise for environmental and economic challenges.

Bacillus subtilis Simultaneously Detoxified Aflatoxin B1 and Zearalenone

The co-occurrence of aflatoxin B1 (AFB1) and zearalenone (ZEN) in grain-based food and animal feed poses significant health risks to humans and animals due to their potent mutagenic, cytotoxic, and carcinogenic properties. Conventional physical and chemical methods are insufficient for effectively detoxifying multiple mycotoxins present in food and feed. In this study, we evaluated the capability of Bacillus subtilis ZJ-2019-1 (B. subtilis ZJ-2019-1) to simultaneously degrade AFB1 and ZEN while optimizing reaction to enhance degradation efficiency. The localization of active ingredients from B. subtilis ZJ-2019-1 was determined using high-performance liquid chromatography (HPLC). Our findings demonstrated that B. subtilis ZJ-2019-1 eliminated 60.88% of AFB1 and 33.18% of ZEN within 72 h at a concentration of 10 mg/L at 37 °C (pH 7.0) and exerted greater activity under alkaline conditions. The autoclaved and boiled supernatants of B. subtilis ZJ-2019-1 exhibited significant enhancement in the degradation of AFB1 and ZEN, achieving degradation rates of 79.85% and 100%, respectively, at a concentration of 1 mg/L within 48 h at 37 °C. Moreover, the crude enzymes from B. subtilis ZJ-2019-1 showed maximum degradation rates for AFB1 (100%) and ZEN (94.29%) within 72 h at 70 °C. Additionally, divalent cations (such as Co2+, Fe2+, Mn2+, and Ni2+) significantly augmented the activity of crude enzymes from B. subtilis ZJ-2019-1 towards mycotoxin degradation. Furthermore, when applied to corn gluten meals, B. subtilis ZJ-2019-1 strain effectively detoxify 66.08% of AFB1 and 22.01% of ZEN, surpassing the efficacy of a commercial detoxification agent on the market (34.17% for AFB1 and 2.28% for ZEN). Collectively, these findings indicated that B. subtilis ZJ-2019-1 is a promising candidate for the simultaneous removal of multiple mycotoxins in food and feed, while addressing health concerns associated with harmful mycotoxins.

Isolation, Molecular Characterization, Optimization and Purification of Amylase Enzyme from Locally Isolated Bacillus Species in Different Regions of Munshiganj, Bangladesh

Amylase is a kind of enzyme that facilitates the breakdown of carbohydrates in the body. This enzyme has many applications in different industries. Generally, in pharmaceutical sector it is used for the treatment of pancreatic disorder, pancreatic enzyme replacement therapy (PERT) and as a digestive aid. In this study, the bacterial strain was isolated from soil samples collected from potato dumpsites in different areas of Munshiganj, Bangladesh. After subculturing on the nutrient agar plate, 31 colonies were obtained, of which 9 isolates were found as amylase producer on starch agar medium. Of these, 2 isolates (MC-04 and MC-15) were selected based on the starch hydrolysis clear zone ratio. The isolate MC-04 showed crude enzyme activity of 2.82 IU/ml and specific enzyme activity of 3.42 IU/mg, and isolate MC-15 showed crude enzyme activity of 3.16 IU/ml and specific enzyme activity of 3.77 IU/mg. The best isolate, MC-15, was then identified by morphology, biochemical and molecular characterization and confirmed by 16S-rRNA gene sequencing. After 16S-rRNA sequencing, the isolate (MC-15) was identified as Bacillus subtilis. This amylase production of this strain had also been optimized under certain conditions such as different incubation periods, pH, temperatures and different carbon sources. We found that the best incubation period was 48 h, the optimum pH-7.0, the optimum temperature at 40°C and 2% starch was considered as the best source of carbon. Finally, the crude amylase enzyme was purified by precipitation with ammonium sulfate, dialysis, and single-step gel filtration chromatography. The enzymatic activity of the purified amylase was found to be 8.91 IU/ml, that was 2.82-fold greater enzymatic activity than the raw enzyme. The experiments confirmed that Bacillus subtilis may be a good source of amylase enzyme for industrial application in Bangladesh. Dhaka Univ. J. Pharm. Sci. 22(2): 147-154, 2023 (December)

Fibrinolytic Protease Production: Impact of Initial pH and Temperature in Solid-State Fermentation by Rhizopus microsporus var. oligosporus FNCC 6010

Background: Fibrinolytic enzyme is one of the cardiovascular disease therapies. Rhizopus microsporus var. oligosporus is microorganism that has been evaluated to produce fibrinolytic protease by fermentation. This study conducted fermentation of helianthi annui semen (sunflower seed) by Rhizopus microsporus var. oligosporus to produce fibrinolytic enzyme. Objective: This study aims to determine the effect of Initial pH and incubation temperature and its optimization in the production of fibrinolytic protease by Rhizopus microsporus var. oligosporus FNCC 6010 in solid-state fermentation on helianthi annui semen (sunflower seed) substrate. Optimum condition was determined by maximum protease and fibrinolytic activity. Method: A crude enzyme of protease fibrinolytic was obtained from the supernatant extract of fermented sunflower seed. Protease activity was measured by the skimmed milk agar (SMA) plate method, and fibrinolytic activity was determined by the fibrin agar plate method. Result: It was found that the starting pH affects both the proteolytic and fibrinolytic activity of enzymes that are produced in fermentation. The starting pH of 5.0 showed higher fibrinolytic and proteolytic activity values compared to the starting pH of 7.0. The incubation temperature 33±1 °C had the higher activity compared to 28±1 °C or 37±1 °C. Conclusion: Initial pH and incubation temperature affect the proteolytic and fibrinolytic activity of crude enzyme extracted from fermented sunflower seed by Rhizopus microsporus var. oligosporus. The optimum condition for producing fibrinolytic protease in the state fermentation method was an initial pH of 5.0 and an incubation temperature of 33±1°C.

Ultraviolet irradiation effect on characteristics of protease enzyme from African catfish (Clarias sp.) intestines

The goal of this study was to investigate the impact of UV-C irradiation on the activity of crude enzymes derived from African Catfish intestines as well the optimum conditions for subsequent autolysis processes. Three key variables were assessed: UV-C exposure time, pH and temperature. The experiments involved the homogenization of intestines in cold distilled water followed by subjecting the mixture into UV-C treatment for exposure times of 0, 10, 20 and 30 minutes. After centrifugation process, the collected supernatant was utilized as the crude enzyme. At various level of temperature (40, 50, 60°C) and pH (6, 7, 8) the activity of crude enzyme was assayed with casein as substrate. The findings revealed the increasing of EA as the UV-C exposure time decreased and pH increased, while the highest EA value was observed at temperature of 50°C. Consequently, the optimal conditions were identified as follows: 0 minutes of UV-C exposure time, pH of 8, and temperature of 50°C. Furthermore, UV-Vis absorption, FTIR, and fluorescence spectroscopy of the crude enzyme after UV-C irradiation induction was studied to investigate the its conformational changes. These additional analyses provided valuable insights into the structural alterations of the crude enzyme caused by the UV-C treatment.

Study on active capacity and detergent application potential of low-temperature alkaline serine protease produced by new strain Exiguobacterium indicum 1.2.3

Microorganisms have long captivated researchers for their potential to produce enzymes with diverse industrial applications. Efficient production of proteases from new strains is crucial as these enzymes play a vital role in breaking down protein bonds, enabling their use in industrial applications. Therefore, a novel Exiguobacterium indicum 1.2.3 was isolated (Istanbul, Turkiye) and characterized in this study. This strain produced alkaline serine protease, which works in lower temperatures (20–40 °C) with casein as a specific substrate. The protease was utterly stable for 3 h at 30 °C. The enzyme was also highly stable in the pH range of 8–11. The optimum activity was obtained at pH 10. The crude enzyme activity was enhanced by various metal ions and retained 147%, 125%, 124%, and 117% of its activity within 1 mM Ca2+, Mn2+, Cu2+, and Mg2+, respectively. The crude enzyme was inactive with phenylmethylsulfonyl fluoride, indicating a serine residue on the active side. The enzyme exhibited a significant proteolytic effect in the presence of surfactants and oxidizing agents. The addition of Tween 80, Triton X-100, and sodium perborate improved enzymatic activity up to 135%, 109%, and 105%, respectively. According to the washing results, the crude enzyme effectively removed the blood on different types of standard pre-stained textiles at 30 °C. In conclusion, Exiguobacterium indicum 1.2.3 is a promising candidate for protease production, with its diverse applications spanning various industrial sectors, particularly detergents.Graphical

CRUDE CELLULASES FROM SUBTERRANEAN TERMITE EXTRACT (Macrotermes gilvus Hagen) FOR SACCHARIFICATION OF LEAF LITTER AND BIOSOFTENING PROCESS IN SKELETON LEAF CRAFTS MAKING

Recently, leaf litter has been used as compost or piled up and burned openly in most regions of Indonesia. This litter has the potential for its lignocellulosic components as the source of sugars. Also, the leave skeleton structure is promising for creative industrial purposes. Besides, termites (Macrotermes gilvus Hagen) can digest lignocellulosic biomass containing high fiber. In this research, termite extract contained lignocellulases and was used in the leaf litter pretreatment process for sugar preparation and leaf litter biosoftening. This study aimed to extract termite enzymes, identify, evaluate the activity of crude enzymes, and characterize the hydrolytic ability of enzymes to degrade lignocellulosic materials in three different leaf litters, i.e., Polyalthia longifolia (PL), Pterocarpus indicus (PI), and Filicium decipiens (FD). The leaf litter biosoftening process was performed in pulp removal for Annona muricata (AM) skeleton leaf crafts making. The results showed that M. gilvus Hagen extract exhibited high activity of endo-β-D-1,4-glucanase (CMCase), moderate activity of avicelase, β-D-1,4-mannanase, β-D-1,4-xylanase, and low β-D-1,4-glucosidase. The optimum pH of CMCase, β-D-1,4-mannanase, and β-D-1,4-glucosidase were 6.5, 5.0, and 6.0, respectively, while the optimum temperatures were 40 °C, 50 °C, and 55 °C, respectively. Cellulases from termite extracts were suitable pretreatment of leaf litter biomass, as evidenced by saccharification activity, with the highest saccharification activity in FD leaf litter. Enzymes were also successfully employed to help remove AM leaf pulp at the volume of 100 mL for 14 days, with the removal efficiency at 98.7%.

Fibrinolytic Protease Activity of Crude Enzyme from Fermented Sunflower (Helianthus annuus) and Common Bean (Phaseolus vulgaris) seeds by Rhizopus microsporus var. oligosporus FNCC 6010 in Solid State Fermentation

In the entire world, cardiovascular diseases (CVDs) are the main cause of death. For the treatment of CVDs, microbial fibrinolytic enzymes are highly regarded as novel therapeutic candidates. This study was purposed to determine the fibrinolytic protease activity produced by fungus source, which is Rhizopus microsporus var. oligosporus FNCC 6010 in fermented sunflower (Helianthus annuus) seed and common bean (Phaseolus vulgaris) seed. Fermentation was carried out by solid-state fermentation method at an initial pH of 5, incubation temperature of 33±1°C, and incubation time of 24 hours. The fermented seed was extracted to obtain supernatant as the crude enzyme. The proteolytic activity assay was done by the skimmed milk agar (SMA) plate method to obtain the proteolytic index, and the fibrinolytic activity assay was conducted by the fibrin-agarose plate method to get the fibrinolytic index. The results show that crude enzymes from fermented H. annuus and P. vulgaris seeds by R. microsporus have fibrinolytic protease activity with proteolytic index 2.64 ± 0.01 and 2.23 ± 0.04, respectively. The fibrinolytic index is 2.40 ± 0.06 and 1.64 ± 0.06, respectively. Therefore, the crude enzyme has the potential to be further researched as a candidate for thrombolytic agents. The purification, characterization, and in-depth research are needed to develop enzymes into preparations for preventing and treating CVDs.

PRODUCTION OPTIMIZATION, PARTIAL PURIFICATION, AND THROMBOLYTIC ACTIVITY EVALUATION OF PROTEASE OF Bacillus cereus HSFI-10

Cardiovascular disease is the primary cause of mortality in the world due to the formation of blood clots or thrombi in blood vessels. Bacterial proteases commonly function as thrombus dissolver agents in the pharmaceutical industry. Bacterial isolate HSFI-10 (Holothuria scabra Fermented Intestine-10) previously isolated from Rusip fermented sea cucumber had demonstrated thrombolytic activity. This study aimed to produce crude protease of HSFI-10 strain at an optimized incubation time and determine the thrombolytic activity of crude and dialysate proteases on A, B, AB, and O blood types. Isolate HSFI-10 was first molecularly identified and found to be Bacillus cereus with a homology level of 99.80% with Bacillus cereus strain ST06. The optimum crude enzyme was obtained after 48-h incubation with an activity of 222.52 U/mL, which increased to 438.84 U/mL after ammonium sulfate precipitation and dialysis. Clot lysis activity of crude enzymes was measured based on the gravimetry method on blood in the ABO system, showing results that ranged from 68.99% to 69.76%, while the dialysate ranged from 81.16% to 82.52%. In conclusion, partial purification of bacterial protease could increase both its specific and thrombolytic activities on human blood in the ABO system, with only 1% activity variability between A, B, AB, and O blood types.

Regulation of Density and Fertilization on Crude Protein Synthesis in Forage Maize in a Semiarid Rain-Fed Area

Density and fertilization mode are the key factors regulating crude protein synthesis in forage maize; however, there is a lack of systematic understanding of the regulation mechanism. Here, the nitrogen/potassium ratio (N/K), free amino acid (AA) content, crude protein synthesis key enzyme activities (nitrate reductase (NR), glutamine synthetase (GS) and glutamic pyruvic transaminase (GPT)) and crude protein content and yield in leaves, stems, and grain of forage maize, as well as the relationships among them, were explored. The results showed that the N/K of DL-40 (60,000 plants per ha−1, 40% N topdressing at large trumpet stage) and DH-50 (75,000 plants per ha−1, 50% N topdressing at large trumpet stage) significantly increased in leaves, stems, and grain, and correspondingly, NR, GS, and GPT activities in leaves, NR activities in stems, NR and GPT activities in grain, and delayed the decline in AA content. After tasseling, for DL-40 and DH-50 the crude protein content increased by 74.1% and 39.8% in leaves, respectively, 19.9% and 25.6% in grain, respectively, and crude protein yield increased by 16.7% and 35.2% in leaves, respectively, and 23.5% and 25.9% in grain, respectively. There were significant quadratic parabolic relationships of NR, GS, and GPT activities with proportion of topdressing. There was a significant relationship of crude protein content with NR activity in leaves, with NR, GS, and GPT activities in stems and with GPT activity in grain. Regulating the key enzyme activity by adjusting the density and fertilization can significantly improve the crude protein yield of forage maize. Treatments DL-40 and DH-50 significantly increased crude protein content and yield by increasing plant N/K, NR activity in leaves, NR activity in stems, and GPT activity in grain, but slowed the decrease in AA content in leaves, stems, and grain.

Optimization of laccase enzyme extraction from spent mushroom waste of Pleurotus florida through ANN-PSO modeling: An ecofriendly and economical approach

The cardinal focus of this study is to optimize the best reaction conditions for maximizing laccase activity from spent mushroom waste (SMW) of Pleurotus florida. Optimization process parameters were studied by the modeling techniques, artificial neural networking (ANN) embedded in particle swarm optimization (PSO), and response surface model (RSM). The best topology of ANN-PSO architecture was obtained on 4-10-1. The R2, IOA, MSE, and MAE values of the ANN model were obtained as 0.98785, 0.9939, 0.0023, and 0.0251 while, that of the RSM model were obtained as 0.74290, 0.9210, 0.0244, and 0.1110 respectively. The higher values of R2, IOA, and lower values of MSE and MAE of the ANN-PSO model depict that ANN-PSO outperformed compared to RSM and also verified the effectiveness of the ANN-PSO model. The ANN-PSO model performance demonstrates the robustness of the technique in optimizing laccase activity in SMW of P. florida. The optimization results revealed that pH 4.5, time 3 h, solid: solution ratio 1:5, and ABTS concentration of 1 mM was optimal for achieving maximum laccase activity at temperature 30 °C. The enzymatic activity of crude laccase enzyme was obtained as 1.185 U ml−1 without loss of enzyme activity. Additionally, crude laccase enzyme was 1.74 fold partially purified, and 83.54% of the enzyme was yielded. Out of all the independent process variables, ABTS and pH had an influence on laccase activity. Therefore, we anticipate that the findings of this investigation will reduce the ambiguity in maximizing laccase activity and ease the screening process. This study also highlights the comparative cost evaluation of crude laccase enzyme extracted from P. florida and commercial enzymes. There is a great potential for the utilization of the laccase enzyme extracted from SMW and using it for the degradation of recalcitrant micropollutants. Thus, SMW promises a cost-effective and sustainable approach leading towards circular economy.

Effect of zinc uptake on alcohol dehydrogenase, protein and mineral contents of hydroponically grown chickpea (Cicer arietinum)

Biofortification of micronutrients is a cost-effective and sustainable approach toward addressing micronutrient malnutrition prevailing across the globe. Understanding Zn uptake dynamics is critical for Zn biofortification of chickpea grains. This study employed the treatment of chickpea with different levels of ZnSO4.7H2O and its effect on growth parameters. 2 µM concentration of Zn + for 12 hrs resulted in significantly higher crude enzyme (Alcohol dehydrogenase) content, enzyme activity and Zn content as compared to all other treatment. Zn content increased significantly with higher Zn application with ZnSO4 concentration with respect to time, giving a maximum increase of 175.22% with 4 µM of Zn. On basis of experimental findings, it is suggested that incorporating 2 µM of Zn in culture solution up to 12 hrs will probably be the best combination of treatment to improve the plant growth, development and quality parameters of chickpea.

Uji Beberapa Jamur Antagonis Terhadap Colletotrichum gloeosporiodes Penyebab Penyakit Busuk Buah Apel Manalagi (Malus sylvestris) Secara In Vitro

Colletotrichum Gloeosporiodes is one of the pathogenic fungi that causes fruit rot disease in apple plants. The control that has been widely used so far is chemical fungicides which cause negative effects, it is necessary to use an alternative control using antagonistic fungi that is safe and environmentally friendly. This study aims to determine the inhibition of several antagonistic fungi against the growth of the pathogen C. gloeosporiodes. The method used was a completely randomized design (CRD) with 4 types of antagonist fungi, 1 control and 5 replications. Antagonism test using dual culture method between antagonistic fungi Trichoderma harzianum, Trichoderma viridae, Trichoderma koningii, and Gliocladium sp. with C. gloeosporiodes. The test results showed that the antagonist fungus that had the highest percentage of inhibition against C. gloeosporiodes was T. harzianum at 87.8% and had a high inhibitory power category with a crude extract chitinase enzyme activity value of 4.30 U/mL and Gliocladium sp. has the lowest percentage of inhibition 64.3%. Based on these results, it can be concluded that T. harzianum has a high potential to control fruit rot disease in apples because it contains the chitinase enzyme which can degrade the cell wall of the pathogenic C. gloeosporiodes.

UTILIZATION OF AGRICULTURAL WASTE AS A SUBSTRATE FOR PRODUCING CELLULASE ENZYME BY ASPERGILLUS NIGER

Fresh Fruit Bunches (FFB) of oil palm, bran, straw, and bagasse are agricultural wastes whose availability is very abundant in Indonesia. The agricultural waste is lignocellulosic waste which still has economic value if further processing is carried out, namely as a substrate in the production of cellulase enzymes. Cellulase enzymes are commonly used in various industries such as food biotechnology, textiles, animal feed, paper, and agriculture to degrade cellulose with its main products, namely glucose, cellobiose, and cellooligosaccharides. In producing cellulase enzymes, it is necessary to have microorganisms that have a high ability to produce enzymes, one of which is Aspergillus niger. The purpose of this study was to determine the activity of crude cellulase enzyme, protein content, and specific activity of cellulase enzyme from agricultural waste which includes FFB, bran, straw, and bagasse. The research methods included preparation of Aspergillus niger culture, delignification, basal medium preparation, cellulase enzyme production, enzyme extraction, crude cellulase enzyme activity test (CMC-ase), lowry method protein content test, and determination of cellulase enzyme specific activity. The study showed that the highest crude cellulase enzyme activity in bran was 26.83 U/ml, the highest protein content in bagasse was 63.42 g/ml, and the highest specific activity of cellulase enzyme in straw was 0.9818 U/ml. The high enzyme activity is influenced by the cellulose content in the material, type of substrate, media, substrate concentration, pH, and temperature.

Extraction,purification and characterization of amylase,protease,lipase of halophilic bacterial isolates from Bordi, India

Extremely halophilic isolates were obtained from mangrove vegetation of west coast Bordi region, Maharashtra India. All six isolates were selected for further investigation on the basis of their potential of producing enzymes amylase, protease and lipases respectively. Production of enzyme from isolates was performed using basal medium with starch, skimmed milk and tributyrin. Ammonium sulphate salt precipitation and dialysis was performed to obtain partially purified crude samples of enzymes. Activity of crude enzymes was determined in unit/ml by using standard methods. Also the effect of pH and temperature on enzyme activity was also determined.

Antifungal Activity and Properties of Crude Enzymes Produced from Cellulomonas fimi JH-1

In this study, bacteria were isolated from rosemary rhizosphere and chitinolytic activity was detected in 0.2% chitin agar medium. Using 16s rRNA sequencing, the isolated bacteria were identified as Cellulomonas fimi. Chitinase, cellulase, and protease activities were found to increase with incubation time. Chitosanase activity was highest on day 1, and gradually declined thereafter. Crude enzyme of C. fimi was prepared and separated by SDS-PAGE and the expression patterns of enzyme activity for chitinase and chitosanase isozymes was studied. Three chitinase isozymes (36, 55, and 66 kDa) and one chitosanase (39 kDa) were detected SDS-PAGE gel combinded with glycol chitin and glycol chitosan, and these chitinase and chitosanase had different molecular weights. Indole acetic acid (IAA) production by C. fimi was analyzed in nutrient broth (NB) media containing different concentrations of tryptophan. IAA production was found to increase with increasing tryptophan concentration. Siderophore production by C. fimi was analyzed in colloidal chitin medium for 0, 1, 3, 5, and 7 d. The production of siderophore increased for 5 d and gradually decreased thereafter. The antifungal activities of C. fimi against G. zeae, F. solani, B. cinerea, F. oxysporum, C. cucumerinum, R. solani, and C. acutatum were investigated in potato dextrose broth medium containing chitin. C. fimi showed the highest antifungal activity against G. zeae.

The Properties of Wet Blue Added Crude Enzyme from Rhizopus oligosporus in the Acid Bating Process

Bating is one of the processes in leather production. This process is generally performed under slightly alkaline conditions, but it can also be done under acidic conditions. Rhizopus oligosporus is categorized as a microorganism that can synthesize acidic enzymes optimally in acidic conditions. This study aimed to evaluate the effect of the bating process using acid protease from R. oligosporus on the leather qualities tanned by chromium sulfate. The study used a completely randomized design where crude enzyme activity (0, 2.5, 5, 7.5, and 10 U mL-1) was a single treatment as an experimental design. Data were analyzed using ANOVA and furtherly tested using Duncan. The results show that the protease synthesized from R. oligosporus is potential to be used as a bating agent. The treatment with 2.5 U mL-1 activity was the best treatment in this study to improve leather (wet blue) properties like shrinkage temperature, ash content, and chrome oxide content. The quality of wet blue produced was better than that produced with conventional treatments. It can be concluded that the enzyme activity of 2.5 U mL-1 could be used as a bating-agent alternative in the acid bating stage.