Screening and isolation of an organic solvent-tolerant bacterium for high-yield production of organic solvent-stable protease

Tang, Xiaoyu; Pan, Yong; Li, S.; He, Bingfang

doi:10.1016/j.biortech.2008.01.030

Cited by 51 publications

(35 citation statements)

References 19 publications

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“…SOMP is highly stable in range of solvents but its stability does not correlate with the log P values. It is in agreement with previous reports [6,26]. SOMP activity was not only stable but also enhanced by octane, heptanes, hexane, toluene, dichloromethane, ethylacetate, and DMSO by various extents, which is highly encouraging.…”

Section: Discussionsupporting

confidence: 93%

An Oxidant- and Organic Solvent-Resistant Alkaline Metalloprotease from Streptomyces olivochromogenes

Simkhada

Cho

Park

et al. 2010

Appl Biochem Biotechnol

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Organic solvent- and detergent-resistant proteases are important from an industrial viewpoint. However, they have been less frequently reported and only few of them are from actinomycetes. A metalloprotease from Streptomyces olivochromogenes (SOMP) was purified by ion exchange with Poros HQ and gel filtration with Sepharose CL-6B. Apparent molecular mass of the enzyme was estimated to be 51 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gelatin zymography. The activity was optimum at pH 7.5 and 50 degrees C and stable between pH 7.0 and 10.0. SOMP was stable below 45 degrees C and Ca(2+) increased its thermostability. Ca(2+) enhanced while Co(2+), Cu(2+), Zn(2+), Mn(2+), and Fe(2+) inhibited the activity. Ethylenediaminetetraacetic acid and ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, but not phenylmethylsulfonyl fluoride, aprotinin, and pefabloc SC, significantly suppressed the activity, suggesting that it might be a metalloprotease. Importantly, it is highly resistant against various detergents, organic solvents, and oxidizing agents, and the activity is enhanced by H(2)O(2). The enzyme could be a novel protease based on its origin and peculiar biochemical properties. It may be useful in biotechnological applications especially for organic solvent-based enzymatic synthesis.

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Section: Discussionsupporting

confidence: 93%

An Oxidant- and Organic Solvent-Resistant Alkaline Metalloprotease from Streptomyces olivochromogenes

Simkhada

Cho

Park

et al. 2010

Appl Biochem Biotechnol

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“…One gram of sample was suspended in 10 ml sterile sea water. After shaking, 5 ml suspension was added into a 250 ml Erlenmeyer flask containing 25 ml of modified Luria-Bertani (MLB) medium (Tang et al, 2008). Toluene was added to the medium at a concentration of 10% (v/v).…”

Section: Enrichment Of Organic Solvent Tolerant Microorganismsmentioning

confidence: 99%

Screening and identification of a novel organic solvent-stable lipase producer

Shu

Fang

et al. 2009

Ann. Microbiol.

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-A strain named DS9 excreting organic solvent-stable lipase was screened and later identified as Bacillus subtilis based on its phenotypes, biochemical test, and 16S rRNA gene sequence. Strain DS9 grows well on the medium with 10% (v/v) organic solvent with log P values equal to or above 2.5. The organic solvent-tolerant lipase excreted by strain DS9 had a wider tolerance for organic solvents. The relative activity of the lipase was above 60% at 37 °C, 200 rpm, 30 min in the present of 25% (v/v) organic solvents such as 1-butanol, hexanol, benzene, and toluene. The lipase was not only stable but also activated by n-hexane, xylene, heptane, isooctane, and n-decane. The optimal pH and temperature were 8.0 and 40 °C, respectively. Both the organic solvent-tolerant microorganism and the organic solvent-stable lipase produced by this strain could be used as a biocatalyst for application in non-aqueous biocatalysis.

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“…The crude protease was quite stable in the presence of 50% solutions of various organic solvents [17]. In this study, we report the purification and characterization of a protease from P. aeruginosa PT121.…”

Section: Introductionmentioning

confidence: 96%

Biochemical Properties and Potential Applications of a Solvent-Stable Protease from the High-Yield Protease Producer Pseudomonas aeruginosa PT121

Tang

Ying

et al. 2009

Appl Biochem Biotechnol

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An organic solvent-stable protease from Pseudomonas aeruginosa PT121 was purified in a single step with 55% recovery by hydrophobic interaction chromatography on a Phenyl Sepharose High Performance matrix. The purified protease was homogenous on SDS-PAGE and had an estimated molecular mass of 33 kDa. The optimal pH and temperature conditions for enzyme activity were 8.0 and 60 degrees C, respectively. The enzyme was classified as a metalloprotease based on its strong inhibition by EDTA and 1,10-phenanthroline and exhibited good stability across a broad pH range (6.0-11.0). The protease was quite stable in the presence of various water-miscible organic solvents. This is a unique property of the protease which makes it an ideal choice for application in aqueous-organic phase organic synthesis including peptides synthesis. The synthetic activity of the protease was tested using N-carbobenzoxy-L-asparagine (Z-Asp) and L-phenylalaninamide (Phe-NH2) as substrate in the presence of various water-miscible organic solvents for aspartame precursor synthesis. The highest yield was obtained in the presence of 50% DMSO (91%). The synthesis rate in the presence of DMSO was also much higher than the rates in the other tested organic solvents, and the initial rates of Z-Asp-Phe-NH2 synthesis in mixtures of various water-miscible organic solvents, with the exception of ethanol, correlated with the yields of Z-Asp-Phe-NH2. Furthermore, the PT121 protease was able to use various carboxyl components (Z-AA) and Phe-NH2 as substrates to catalyze the syntheses of the dipeptides, indicating that this protease has a broad specificity for carboxylic acid residue.

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Screening and isolation of an organic solvent-tolerant bacterium for high-yield production of organic solvent-stable protease

Cited by 51 publications

References 19 publications

An Oxidant- and Organic Solvent-Resistant Alkaline Metalloprotease from Streptomyces olivochromogenes

An Oxidant- and Organic Solvent-Resistant Alkaline Metalloprotease from Streptomyces olivochromogenes

Screening and identification of a novel organic solvent-stable lipase producer

Biochemical Properties and Potential Applications of a Solvent-Stable Protease from the High-Yield Protease Producer Pseudomonas aeruginosa PT121

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