Lipase production bacterial isolate was isolated from soil of service station and identified as Bacillus methylotrophicus PS3 by 16SrRNA with accession number |LN999829.1|. Lipase enzyme was purified by sequential methods of ammonium sulfate precipitation and Sephadex G-100 gel column chromatography. The molecular weight of purified enzyme was 31.40 kDa on SDS-PAGE. This purification procedure resulted in 2.90-fold purification of lipase with a 24.10% final yield. The purified lipase presented maximal hydrolytic activity at a temperature of 55 °C, and pH of 7.0. Lipase activity was stimulated by Triton X-100 and SDS with Mg2+ and Ca2+ metals employ a positive effect and outlast its stable in organic solvent i.e. methanol and ethanol.
Microbial origin polysaccharides have gained popularity due to lesser toxicity, better degradability and selectivity as compared to their synthetic counterparts and can be used as emulsifier, stabilizer, thickener, texturizer, flocculating and gelling agent. Here main emphasis on exopolysaccharide production from potential lactic acid bacteria that has GRAS status. This work was aimed at isolating, purifying and characterizing an extracellular polysaccharide (EPS) produced by a foodgrade lactic acid bacteria Lactobacillus paraplantarum KM1. L. paraplantarum KM1 was isolated from human milk and identified by conventional and molecular techniques. The 16S rRNA sequence of the isolate was registered in National Centre for Biotechnology Information (NCBI) under accession number KX671558. L. paraplantarum KM1 was found to produce EPSs in lactose containing MRS medium, and the maximum yield (47.4 mg/ml) was achieved after 32-h incubation. As evident from TLC and HPLC analyses, the polysaccharide was found to be a heteropolymer-containing glucose, galactose and mannose as main sugars. Different oligosaccharides namely hexoses were obtained after partial hydrolysis of the polymer using MALDI-ToF-MS. The total molecular weight of all polysaccharides present was 348.7 kDa with 100 °C thermal stability as well as water soluble in nature. Cell cytotoxicity revealed that the purified EPS was safe for consumption; thus, it can be used in various food industries as emulsifying and texture agent. The present study highlighted that exopolysaccharides could be harnessed to improve food products in terms of texture, emulsifying agents, pharmaceutical industry (antioxidants, antitumour, anti-inflammatory and antiviral agents) and as safety purposes.
A novel strain Rhizopus sp. C4 was isolated from compost for the production of pectinase. Cultivation of Rhizopus sp. C4 on orange peel substrate under various solid-state fermentation (SSF) conditions was evaluated for pectinase yield along with the enzyme activity profile as a potential, low-cost alternative to submerged-liquid fermentation. Response surface methodology (RSM) was employed to optimize various environmental parameters for pectinase production. Various parameters, namely temperature, moisture and incubation days, were studied statistically for a total of 20 runs using central composite design. The highest yield of the enzyme, i.e., 11.63 IU/mL, was obtained from 1:3.5 moisture ratios in 7 days at 30°C. The study demonstrated that optimization through RSM could improve the enzymatic characteristics and yield of the enzyme.
6 September 2017 in Biocatalysis and Biotransformation
Apple pomace as a substrate for bioethanol production is interesting due to its abundance and sustainable availability in varied states like Himachal Pradesh (H.P.), Jammu and Kashmir, Uttarakhand and Arunachal Pradesh, India. In the current study, apple pomace which is the main fruit industrial waste of H.P. was evaluated as feedstock for bioethanol production by the process of enzymatic saccharification using multiple carbohydrases. Microwave pretreatment of the apple pomace resulted in the efficient removal of lignin and crystalline structure of cellulose fibre. The enzymatic saccharification of the pretreated biomass was done by optimizing parameters for maximal saccharification leads to production of 27.50 mg/g of reduce, ng sugar. An enhanced ethanol yield of 44.46 g/l and fermentation efficiency of 58% by immobilized co-culture of Saccharomyces cerevisiae MTCC 3089 and Scheffersomyces stipitis NCIM 3498 under SHF as compared to fermentation performed with free yeast cells, i.e. 34.46 g/l of ethanol and 45% of fermentation efficiency.
1 June 2018 in Journal of Genetic Engineering and Biotechnology
The brown rot fungus Rhizopus delemar F2 was shown to produce extracellular thermostable and multiple carbohydrase enzymes. The potential of Rhizopus delemar F2 in utilizing apple pomace under solid state fermentation (SSF) is the purpose of the study. Solid state fermentation (SSF) is a very effective technique opposed to submerged fermentation in various aspects. Enhanced production of multiple carbohydrases 18.20 U g-1 of cellulose, 158.30 U g-1 of xylanase, 61.50 U g-1 of pectinase and amylase 21.03 U g-1 was released by microwave pretreatment of apple pomace at 450 W for 1 min and then by incubation the culture thus obtained at 30 °C for 6 days with moisture content of 1:4.5. Apple pomace can serve as a potential source of raw material for the production of multiple carbohydrases. Besides, it can find great commercial significance in production of bioethanol and various industries like textile, fruit juice, paper and pulp industry.