Ulhas Patil

Production of Alkaline Protease by Solvent-Tolerant Alkaliphilic Bacillus circulans MTCC 7942 Isolated from Hydrocarbon Contaminated Habitat: Process Parameters Optimization

In the present investigation, a newly isolated organic solvent-tolerant and alkaliphilic bacterial strain was reported from a hydrocarbon (gasoline and diesel) contaminated soil collected from the petrol station, Shirpur (India). The strain was identified as Bacillus circulans MTCC 7942, based on phenotype, biochemical, and phylogenetic analysis of 16S rRNA gene sequence. The capability of Bacillus circulans to secrete an extracellular, thermostable, alkaline protease and grow in the presence of organic solvents was explored. Bacillus circulans produced maximum alkaline protease (412 U/mL) in optimized medium (g/L): soybean meal, 15; starch, 10; KH2PO4, 1; MgSO4·7H2O, 0.05; CaCl2, 1; Na2CO3, 8; pH 10.0 at 37°C and 100 rpm. The competence of strain to grow in various organic solvents—n-octane, dodecane, n-decane, N,N-dimethylformamide, n-hexane, and dimethyl sulfoxide, establishes its potential as solvent-stable protease source for the possible applications in nonaqueous reactions and fine chemical synthesis.

Detergent-compatible, organic solvent-tolerant alkaline protease from Bacillus circulans MTCC 7942: Purification and characterization.

ABSTRACT Proteases are now recognized as the most indispensable industrial biocatalyst owing to their diverse microbial sources and innovative applications. In the present investigation, a thermostable, organic solvent-tolerant, alkaline serine protease from Bacillus circulans MTCC 7942, was purified and characterized. The protease was purified to 37-fold by a three-step purification scheme with 39% recovery. The optimum pH and temperature for protease was 10 and 60°C, respectively. The apparent molecular mass of the purified enzyme was 43 kD as revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The Km and Vmax values using casein-substrate were 3.1 mg/mL and 1.8 µmol/min, respectively. The protease remained stable in the presence of organic solvents with higher (>3.2) log P value (cyclohexane, n-octane, n-hexadecane, n-decane, and n-dodecane), as compared to organic solvents with lower (<3.2) log P value (acetone, butanol, benzene, chloroform, toluene). Remarkably, the protease showed profound stability even in the presence of organic solvents with less log P values (glycerol, dimethyl sulfate [DMSO], p-xylene), indicating the possibility of nonaqueous enzymatic applications. Also, protease activity was improved in the presence of metal ions (Ca2+, Mg2+, Mn2+); enhanced by biosurfactants; hardly affected by bleaching agents, oxidizing agents, and chemical surfactants; and stable in commercial detergents. In addition, a protease–detergent formulation effectively washed out egg and blood stains as compared to detergent alone. The protease was suitable for various commercial applications like processing of gelatinous film and as a compatible additive to detergent formulation with its operative utility in hard water.

Ultrasound-assisted improvements in biocatalytic activity and production of organic-solvent stable protease from Bacillus circulans MTCC 7942

This study reports the ultrasound-assisted improvements in biocatalytic activity and production of protease from organic solvent-tolerant, alkaliphilic Bacillus circulans MTCC 7942. The strain withstands 10min ultrasound treatment (33kHz); this treatment promotes the growth of bacterium. The protease production was improved by 1.5-fold on multiple ultrasound treatment at 0, 6, 12h intervals. Although the substrate casein remains unaffected by ultrasound treatment (33kHz, 10min); the treatment had boosted the enzyme activity of purified protease. Also, the ultrasound treated purified protease showed enhancement in enzyme activity in presence of n-dodecane, n-octane, n-hexadecane, n-decane, xylene, butanol, toluene, pyridine and chloroform.

Operative utility of salt-stable proteases of halophilic and halotolerant bacteria in the biotechnology sector

Proteases are universal in existence in all organisms and are needed for metabolic activities of the cell. Also, proteases are the paramount hydrolytic enzymes widely used in industrial sector accounting ~65% of the aggregate worldwide enzyme market. The market revenue of global protease sale is estimated to be

First report of leaf spot disease caused by Corynespora torulosa MCC-1368 on cotton plant from India

2.21 billion by 2021. Currently, a widening expectation of biotechnology sector necessitates the availability of robust protease with industrially suitable operative features. The commercially-compatible protease should show stability and altered specificity in organic solvents, and be economically as well as ecologically sustainable. The proteolytic halophilic and halotolerant microorganisms are a novel source of salt-stable proteases. Beside stable in the presence of significant amount of salt, the halo-proteases could occasionally display the polyextremophilic attributes like tolerance to alkaline pH, high temperature, and organic solvent tolerance etc. This review describes - halophilic/halotolerant proteolytic microorganisms as source of salt-stable protease, mechanisms of high salt tolerance, various physical and nutritional parameters affecting the protease production, purification strategies for protease, characteristics of salt-stable proteases, and commercial significance of salt-stable proteases. The study revises the current status of the research on salt-stable proteases obtained from extremophile with their operative utility in the biotechnology field.

Medicinal Plants as a Source of Therapeutic Agents against HIV Infection

Abstract In present study, the leaf spot disease of cotton plant emerged in the North Maharashtra region of India was reported. The fungal phytopathogen associated with inducing the leaf spot disease symptoms was isolated and characterised. The isolated fungus was identified as Corynespora torulosa (Deposition accession number, MCC-1368; Genbank accession no. MF462072) based on morphological and cultural characteristics and molecular analysis of ITS region. The pathogenicity of fungal phytopathogen was verified by Koch’s postulates. To our knowledge, this is the first report of incidence of leaf spot disease caused by Corynespora torulosa on cotton plant.

Purification and characterization of solvent‐tolerant, thermostable, alkaline metalloprotease from alkalophilic Pseudomonas aeruginosa MTCC 7926

Many plants have been tested for anti-HIV activity. The present review covers the plants reported to be active against HIV.