Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli

被引：36

作者：

Ribitsch, D. ^{[1
]}

Heumann, S. ^{[1
]}

Karl, W. ^{[1
]}

Gerlach, J. ^{[1
]}

Leber, R. ^{[1
]}

Birner-Gruenberger, R. ^{[1
,5
]}

Gruber, K. ^{[1
,4
]}

Eiteljoerg, I. ^{[1
]}

Remler, P. ^{[1
]}

Siegert, P. ^{[6
]}

Lange, J. ^{[6
]}

Maurer, K. H. ^{[6
]}

Berg, G. ^{[1
,3
]}

Guebitz, G. M. ^{[1
,3
]}

Schwab, H. ^{[1
,2
]}

机构：

[1] ACIB GmbH, A-8010 Graz, Austria

[2] Graz Univ Technol, Inst Mol Biotechnol, A-8010 Graz, Austria

[3] Graz Univ Technol, Inst Environm Biotechnol, A-8010 Graz, Austria

[4] Graz Univ, Inst Mol Biosci, A-8010 Graz, Austria

[5] Med Univ Graz, Med Res Ctr, Prote Core Facil, A-8010 Graz, Austria

[6] Henkel AG & Co KGaA, D-40191 Dusseldorf, Germany

来源：

JOURNAL OF BIOTECHNOLOGY | 2012年 / 157卷 / 01期

关键词：

Detergent protease; Stenotrophomonas maltophilia; Alginate beads; BIOLOGICAL-CONTROL; PYTHIUM-ULTIMUM; PROTEIN; BACTERIA;

D O I：

10.1016/j.jbiotec.2011.09.025

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli. Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3. From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45 degrees C. Specific activity of StmPr2 determined with suc-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide as the substrate was 17 +/- 2 U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：140 / 147

页数：8

共 27 条

[11] Bacterial alkaline proteases: molecular approaches and industrial applications [J].