Browse
Submit Data
Databases
API
Help

Dataset Information

10 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

A statistical approach for optimization of alkaline lipase production by ascidian associated-Halobacillus trueperi RSK CAS9.

ABSTRACT: A marine ascidian-associated bacterium, Halobacillus trueperi RSK CAS9, was optimized for lipase production by response surface methodology using marine waste as substrate. The central composite design was employed, and the optimal medium constituents for maximum lipase production (1355.81 U/ml) were determined to be tuna powder (14.58 g/l), olive oil (5.05 ml/l); NaCl (72.42 g/l), temperature (45 °C) and pH 9.0. An alkaline lipase was purified to 8.46 fold with 1193.59 U mg-1 specific activities with the molecular weight of 44 kDa. The activity was substantially inhibited by EDTA and PMSF, indicating that it was a metalloenzyme serine residue which was essential for catalytic activity. Thus, lipase production by microbial conversion of marine fish wastes in this study suggested its potential utilization for the production of high value products.

SUBMITTER: Sathishkumar R

PROVIDER: S-EPMC4980739 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Json Xml

Publications

A statistical approach for optimization of alkaline lipase production by ascidian associated-Halobacillus trueperi RSK CAS9.

Sathishkumar Ramamoorthy R Ananthan Gnanakkan G Iyappan Kathirvel K Stalin Chinnathambi C

Biotechnology reports (Amsterdam, Netherlands) 20150916

A marine ascidian-associated bacterium, Halobacillus trueperi RSK CAS9, was optimized for lipase production by response surface methodology using marine waste as substrate. The central composite design was employed, and the optimal medium constituents for maximum lipase production (1355.81 U/ml) were determined to be tuna powder (14.58 g/l), olive oil (5.05 ml/l); NaCl (72.42 g/l), temperature (45 °C) and pH 9.0. An alkaline lipase was purified to 8.46 fold with 1193.59 U mg-1 ...[more]

PMID: 28352574

Similar Datasets

OPTIMIZATION OF ALKALINE ?-AMYLASE PRODUCTION BY THERMOPHILIC BACILL US SUBTILIS.

Project description:Starch-degrading amylase enzyme is important in biotechnological applications as food, fermentation, textile, paper and pharmaceutical purposes. The aim of current study to isolate alkaline thermostable ?-amylase bacteria and then study the composition of medium and culture conditions to optimize cells growth and a-amylase production.Thermophilic amylase producing bacterium was isolated from local hot water-springs in Gazan city Saudi Arabia.Phylogenetic analysis of 16 S rRNA sequence for the strain revealed that the strain have the same sequence of Bacillus subtilis. Maximum amylase production was observed, when B. subtilis cultured in medium containing starch at concentration 0.5%, and 10 g/L peptones as nitrogen source at pH 8.5 in when it was incubated for 48 h at 45°C.An amylase-producing bacterium were isolated from hot-spring water and was identified as B. subtilis. Amylase produced from B.subtilis had optimum temperature 45°C and pH 8.5 in shaking media.

| S-EPMC5411881 | biostudies-literature

Optimization of polygalacturonase production from a newly isolated Thalassospira frigidphilosprofundus to use in pectin hydrolysis: statistical approach.

Project description:The present study deals with the production of cold active polygalacturonase (PGase) by submerged fermentation using Thalassospira frigidphilosprofundus, a novel species isolated from deep waters of Bay of Bengal. Nonlinear models were applied to optimize the medium components for enhanced production of PGase. Taguchi orthogonal array design was adopted to evaluate the factors influencing the yield of PGase, followed by the central composite design (CCD) of response surface methodology (RSM) to identify the optimum concentrations of the key factors responsible for PGase production. Data obtained from the above mentioned statistical experimental design was used for final optimization study by linking the artificial neural network and genetic algorithm (ANN-GA). Using ANN-GA hybrid model, the maximum PGase activity (32.54 U/mL) was achieved at the optimized concentrations of medium components. In a comparison between the optimal output of RSM and ANN-GA hybrid, the latter favored the production of PGase. In addition, the study also focused on the determination of factors responsible for pectin hydrolysis by crude pectinase extracted from T. frigidphilosprofundus through the central composite design. Results indicated 80% degradation of pectin in banana fiber at 20 °C in 120 min, suggesting the scope of cold active PGase usage in the treatment of raw banana fibers.

| S-EPMC3881631 | biostudies-literature

Efficient heterologous expression of an alkaline lipase and its application in hydrolytic production of free astaxanthin.

Project description:Background:Astaxanthin, a naturally occurring carotenoid pigment molecule, displays strong antioxidant, anti-cancer, and immunity-enhancing properties, and is often utilized in food, biomedical, cosmetic, and other industries. Free astaxanthin has better solubility than astaxanthin esters (Ast-E), and is a useful auxiliary ingredient in health foods and medicines. Our goal was to establish an improved enzymatic method for preparation of free astaxanthin from natural sources (e.g., the microalga Haematococcus pluvialis), to expand the potential applications of free astaxanthin. Results:The alkaline lipase gene proalip and its propeptide were cloned and successfully fusion-expressed in Pichia pastoris X-33. The recombinant lipase was termed Lipase-YH. Through optimization of culture conditions (medium formulation, pH, added methanol concentration), cell growth (OD600) and secreted enzyme activity respectively reached to 280 and 2050 U/mL in a 50-L autofermentor. Activity of Lipase-YH enzyme powder was about 40,000 U/g. Hydrolysis of Ast-E (extracted from H. pluvialis) by Lipase-YH occurred in aqueous phase, and reaction conditions were optimized based on emulsification method and enzyme/substrate ratio. The highest enzymatic reaction rate was observed for substrate concentration 200 ?g/mL, with maximal free astaxanthin yield (80%) at 1 h, and maximal Ast-E hydrolysis rate 96%, as confirmed by TLC, HPLC, and mass spectroscopy. Conclusion:A novel, efficient enzymatic process was developed for production of free astaxanthin through hydrolysis of Ast-E. Lipase activity was enhanced, and production cost was greatly reduced. The unique structure of free astaxanthin allows linkage to various functional compounds, which will facilitate development of novel pharmaceutical and food products in future studies.

| S-EPMC6020301 | biostudies-literature

Statistical optimization of production conditions of ?-glucosidase from Bacillus stratosphericus strain SG9.

Project description:The present study illustrates the optimization and characterization of ?-glucosidase from a bacterial isolate, strain SG9. Sixty-eight different variables were first screened by one factor at a time method. The screened variable optimization was then performed by Plackett-Burman design followed by Box-Behnken response surface methodology. Thirty-one variables were screened, of which five variables were found to be significant. Box-Behnken design was then performed using the most significant variables, viz., esculin, K2HPO4 and MgSO4. The maximum enzyme activity was observed with an optimal medium composition of esculin (1.9 g/L), K2HPO4 (0. 5 g/L) and MgSO4 (0.3 g/L) with a predicted value of 3392.01 IU. The maximum ?-glucosidase production achieved was 3340 IU. The bacterial strain was identified by 16S rRNA gene sequence and biochemical characterization. The strain was identified as Bacillus stratosphericus and is a first report of its kind.

| S-EPMC5496939 | biostudies-literature

Optimization of a protease extraction using a statistical approach for the production of an alternative meat tenderizer from Manihot esculenta roots.

Project description:Abstract:Meat tenderness is the most important criterion in food quality because it strongly influences the consumer's satisfaction. Tenderness generally depends on connective tissue and sarcomere length of muscle. One of the effective methods for meat tenderizing is protease treatment. In this study, Manihot esculenta root was chosen as a protease source due to its skin blistering effect, suggesting the presence of strong proteolytic activity. The extraction of the crude protease was optimized by using response surface methodology (RSM) with four independent variables, which were pH (X1), CaCl2 (X2), Triton X-100 (X3) and 2-mercaptoethanol (X4). Based on the RSM model, all the independent variables were significant and the optimum extraction conditions were pH 9, 3.24 mM CaCl2, 4.12% Triton X-100 and 6.32 mM 2-mercaptoethanol. Tukey's test results showed that the difference between the expected and experimental protease activity value was 0.05%. A reduction of meat firmness was observed when samples treated with enzyme were compared with a control by using a texture analyser. Electrophoretic patterns also showed extensive proteolysis and a reduction of intensity and number of the protein bands in the treated sample. SEM clearly revealed the degradation of muscle fibres and connective tissue of meat treated with crude protease. Graphic abstract:

| S-EPMC7316877 | biostudies-literature

Production and purification of an alkaline lipase from Bacillus sp. for enantioselective resolution of (±)-Ketoprofen butyl ester.

Project description:The present study was conducted to purify lipase from indigenous Bacillus subtilis strain Kakrayal_1 (BSK-L) for enantioselective resolution of racemic-ketoprofen. The production of lipase (BSK-L) was optimized using Plackett-Burman and central composite design of response surface methodology (RSM). The optimized media containing olive oil (3.5%), MnSO4 (8 mM), CaCl2 (5 mM), peptone (20 g/l), pH (8), agitation (180 rpm) and temperature (37 °C) resulted in maximum lipase production of 7500 U/g of cell biomass. The lipase was purified using sequential method to an overall purification fold of 13% with 20% recovery, 882 U/mg specific activity and a molecular weight of 45 kDa. Optimal pH and temperature of purified lipase were found to be 8 and 37 °C, respectively. Furthermore, BSK-L displayed good stability with various organic solvents, surfactants and metal ions. K m and V max values of lipase were observed to be 2.2 mM and 6.67 mmoles of product formed/min/mg, respectively. The racemic ketoprofen butyl ester was hydrolyzed using lipase with 49% conversion efficiency and 69% enantiomeric excess (ee) which was superior to the commercially procured lipase (Candida antarctica lipase). Thus, this enzyme could be considered as a promising candidate for the pharmaceutical industry.

| S-EPMC6242800 | biostudies-literature

Optimization of alkaline protease production by rational deletion of sporulation related genes in Bacillus licheniformis.

Project description:BackgroundOur laboratory has constructed a Bacillus licheniformis strain that secretes alkaline protease (AprE) with excellent enzymatic properties. B. licheniformis is generally regarded as safe and has a high industrial exoenzyme secretion capacity, but the host retains some undomesticated characteristic that increase its competitiveness and survival, such as spore-formation, which increases the requirements and difficulties in industrial operations (e.g. sterilization and enzyme activity control). Furthermore, the influence of sporulation on alkaline protease production in B. licheniformis has not been elucidated in detail.ResultA series of asporogenic variants of the parent strain were constructed by individually knocking out the master regulator genes (spo0A, sigF and sigE) involved in sporulation. Most of the variants formed abortively disporic cells characterized by asymmetric septa at the poles and unable to survive incubation at 75 °C for 10 min. Two of them (ΔsigF and ΔsigE) exhibited superior characteristics in protease production, especially improving the expression of the aprE gene. Under the currently used fermentation conditions, the vegetative production phase of ΔsigF can be prolonged to 72 h, and the highest protease production of ΔsigF reached 29,494 ± 1053 U/mL, which was about 19.7% higher than that of the wild-type strain.ConclusionWe first constructed three key sporulation-deficient strain to investigate the effect of sporulation on alkaline protease synthesis. The sigF mutant retained important industrial properties such as facilitating the sterilization process, a prolonged stable phase of enzyme production and slower decreasing trend, which will be superior in energy conservation, simpler operations and target product controlling effect. In summary, the work provides a useful industrial host with preferable characteristics and a novel strategy to enhance the production of protease.

| S-EPMC6657089 | biostudies-literature

Combining genotype improvement and statistical media optimization for isoprenoid production in E. coli.

Project description:Isoprenoids are a large and diverse class of compounds that includes many high value natural products and are thus in great demand. To meet the increasing demand for isoprenoid compounds, metabolic engineering of microbes has been used to produce isoprenoids in an economical and sustainable manner. To achieve high isoprenoid yields using this technology, the availability of metabolic precursors feeding the deoxyxylulose phosphate (DXP) pathway, responsible for isoprenoid biosynthesis, has to be optimized. In this study, phosphoenolpyruvate, a vital DXP pathway precursor, was enriched by deleting the genes encoding the carbohydrate phosphotransferase system (PTS) in E. coli. Production of lycopene (a C40 isoprenoid) was maximized by optimizing growth medium and culture conditions. In optimized conditions, the lycopene yield from PTS mutant was seven fold higher than that obtained from the wild type strain. This resulted in the highest reported specific yield of lycopene produced from the DXP pathway in E. coli to date (20,000 µg/g dry cell weight). Both the copy number of the plasmid encoding the lycopene biosynthetic genes and the expression were found to be increased in the optimized media. Deletion of PTS together with a similar optimization strategy was also successful in enhancing the production of amorpha-1,4-diene, a distinct C15 isoprenoid, suggesting that the approaches developed herein can be generally applied to optimize production of other isoprenoids.

| S-EPMC3790805 | biostudies-literature

Statistical-based optimization and scale-up of siderophore production process on laboratory bioreactor.

Project description:We report the enhanced production of siderophore in succinate medium by applying two-stage statistical approach, i.e., Plackett-Burman design and response surface methodology (RSM) using central composite design (CCD). In the first stage of optimization, out of 11 variable components of succinate medium, succinic acid, pH and temperature were found as significant components that influenced the siderophore production in Pseudomonas aeruginosa RZS9. The second stage of RSM using CCD consisted of optimizing the concentrations of the variables. Here, 0.49 g/100 ml concentration of succinic acid, pH 7.08 and temperature of 27.80 °C yielded the maximum (68.41 %) siderophore units. All the significant components exhibited quadratic effect on siderophore production. The F value of 28.63, multiple correlation coefficient (R 2) of 0.9626, percent coefficient of variation of 8.81 values indicated that the model was significant and that the experimental data was satisfactorily adjusted to the quadratic model. During validation of these experiments, 6.10 % increase in siderophore yield was obtained. Scale-up of this protocol optimized at shake flask level up to 5 L-capacity reactor further enhanced the siderophore yield. We claim it to be the first report on statistical optimization of siderophore production by P. aeruginosa RZS9.

| S-EPMC4754294 | biostudies-literature

Efficient Heterologous Production of Rhizopus oryzae Lipase via Optimization of Multiple Expression-Related Helper Proteins.

Project description:This study is dedicated to efficiently produce Rhizopus oryzae lipase (ROL) by optimizing the expression of multiple expression-related helper proteins in Pichia pastoris. A series of engineered strains harboring different copy numbers of the ROL gene and different copies of the chaperone Pdi gene were first constructed to examine the influence of Pdi gene copy number on ROL production. The results showed that multiple copies of Pdi gene did not significantly improve ROL expression. Then, the effect of the co-overexpression of 10 expression-related helper proteins on ROL secretion was investigated by screening 20 colonies of each transformants. The data from shaking-flask fermentation suggested that Ssa4, Bmh2, Sso2, Pdi, Bip, Hac1, and VHb had positive effects on ROL expression. Subsequently, Ssa4, Bmh2, and Sso2, which all participate in vesicular trafficking and strongly promote ROL expression, were combined to further improve ROL production level. ROL activity of the screened strain GS115/5ROL-Ssa4-Sso2-Bmh2 4# attained 5230 U/mL. Furthermore, when the helper proteins Pdi, Bip, Hac1, and VHb were individually co-expressed with ROL in the strain GS115/5ROL-Ssa4-Sso2-Bmh2 4#, lipase activity increased to 5650 U/mL in the strain GS115/5ROL-Ssa4-Sso2-Bmh2-VHb 9#. Additionally, the maximum ROL activity of 41,700 U/mL was achieved in a 3 L bioreactor for high-density fermentation via a sorbitol?methanol co-feeding strategy, reaching almost twofold the value of the initial strain GS115/pAO?-5ROL 11#. Thus, the strategies in this study significantly increased ROL expression level, which is of great potential for the large-scale production of ROL in P. pastoris.

| S-EPMC6274836 | biostudies-literature