Purification and characterization of extracellular alkaline serine protease from Stenotrophomonas maltophilia strain S-1

被引：36

作者：

Miyaji, T

Otta, Y

Shibata, T

Mitsui, K

Nakagawa, T

Watanabe, T

Niimura, Y

Tomizuka, N

机构：

[1] Tokyo Univ Agr, Fac Bioind, Dept Food Sci & Technol, Abashiri, Hokkaido 0992493, Japan

[2] Hokkaido Sugar Co Ltd, Kitami, Hokkaido, Japan

[3] Tokyo Univ Agr, Fac Appl Biosci, Dept Biosci, Setagaya Ku, Tokyo, Japan

来源：

LETTERS IN APPLIED MICROBIOLOGY | 2005年 / 41卷 / 03期

关键词：

alkaline protease; extracellular protease; serine protease; Stenotrophomonas maltophilia; zein;

D O I：

10.1111/j.1472-765X.2005.01750.x

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：

Aims: The present study was conducted by screening soil bacteria in an attempt to isolate a bacterium that produced extracellular alkaline protease, and for purification and characterization of the protease. Methods and Results: Soil bacteria were screened by growth on casein as the sole carbon source. Characterization of a strain isolated from soil of Abashiri, Japan indicated a taxonomic affiliation to Stenotrophomonas maltophilia, and was named S-1 strain. The purified S-1 protease, designed S. maltophilia Protease-1 (SmP-1), exhibited an optimal pH of 12.0, optimal reaction temperature of 50 degrees C and a molecular mass of approximately 40 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The cleavage sites of the oxidized-insulin B chain by SmP-1 were identified as Leu6-Cys7, Cys7-Gly8, Tyr16-Leu17 and Leu17-Val18. The N-terminal amino acid sequence of the purified alkaline protease was determined as NH2-SASAPMVSGVAALVLE. Conclusions: A novel extracellular alkaline serine protease was isolated from S. maltophilia strain S-1. The optimal pH of the proteolytic activity was pH 12.0. Significance and Impact of the Study: The extremely high optimal pH and heat stability of the alkaline serine protease SmP-1 might make it widely applicable to food and other industries.

引用

页码：253 / 257

页数：5

共 17 条

[1] The genome sequence of Drosophila melanogaster [J].

Adams, MD ;

Celniker, SE ;

Holt, RA ;

Evans, CA ;

Gocayne, JD ;

Amanatides, PG ;

Scherer, SE ;

Li, PW ;

Hoskins, RA ;

Galle, RF ;

George, RA ;

Lewis, SE ;

Richards, S ;

Ashburner, M ;

Henderson, SN ;

Sutton, GG ;

Wortman, JR ;

Yandell, MD ;

Zhang, Q ;

Chen, LX ;

Brandon, RC ;

Rogers, YHC ;

Blazej, RG ;

Champe, M ;

Pfeiffer, BD ;

Wan, KH ;

Doyle, C ;

Baxter, EG ;

Helt, G ;

Nelson, CR ;

Miklos, GLG ;

Abril, JF ;

Agbayani, A ;

An, HJ ;

Andrews-Pfannkoch, C ;

Baldwin, D ;

Ballew, RM ;

Basu, A ;

Baxendale, J ;

Bayraktaroglu, L ;

Beasley, EM ;

Beeson, KY ;

Benos, PV ;

Berman, BP ;

Bhandari, D ;

Bolshakov, S ;

Borkova, D ;

Botchan, MR ;

Bouck, J ;

Brokstein, P .

SCIENCE, 2000, 287 (5461) :2185-2195

[2]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[3] Subunit cleavage of mosquito pro-vitellogenin by a subtilisin-like convertase [J].

Chen, JS ;

Raikhel, AS .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (12) :6186-6190

[4] Comparison of the genomes of two Xanthomonas pathogens with differing host specificities [J].

A. C. R. da Silva ;

J. A. Ferro ;

F. C. Reinach ;

C. S. Farah ;

L. R. Furlan ;

R. B. Quaggio ;

C. B. Monteiro-Vitorello ;

M. A. Van Sluys ;

N. F. Almeida ;

L. M. C. Alves ;

A. M. do Amaral ;

M. C. Bertolini ;

L. E. A. Camargo ;

G. Camarotte ;

F. Cannavan ;

J. Cardozo ;

F. Chambergo ;

L. P. Ciapina ;

R. M. B. Cicarelli ;

L. L. Coutinho ;

J. R. Cursino-Santos ;

H. El-Dorry ;

J. B. Faria ;

A. J. S. Ferreira ;

R. C. C. Ferreira ;

M. I. T. Ferro ;

E. F. Formighieri ;

M. C. Franco ;

C. C. Greggio ;

A. Gruber ;

A. M. Katsuyama ;

L. T. Kishi ;

R. P. Leite ;

E. G. M. Lemos ;

M. V. F. Lemos ;

E. C. Locali ;

M. A. Machado ;

A. M. B. N. Madeira ;

N. M. Martinez-Rossi ;

E. C. Martins ;

J. Meidanis ;

C. F. M. Menck ;

C. Y. Miyaki ;

D. H. Moon ;

L. M. Moreira ;

M. T. M. Novo ;

V. K. Okura ;

M. C. Oliveira ;

V. R. Oliveira ;

H. A. Pereira .

Nature, 2002, 417 (6887) :459-463

[5] Purification and characterization of an alkaline serine endopeptidase from a feather-degrading Xanthomonas maltophilia strain [J].

De Toni, CH ;

Richter, MF ;

Chagas, JR ;

Henriques, JAP ;

Termignoni, C .

CANADIAN JOURNAL OF MICROBIOLOGY, 2002, 48 (04) :342-348

[6] ISOLATION AND CHARACTERIZATION OF AN EXTRACELLULAR PROTEINASE PRODUCED BY A SOIL STRAIN OF XANTHOMONAS-MALTOPHILIA [J].

DEBETTE, J .

CURRENT MICROBIOLOGY, 1991, 22 (02) :85-90

[7]

Kalisz H M, 1988, Adv Biochem Eng Biotechnol, V36, P1

[8] Microbial alkaline proteases: From a bioindustrial viewpoint [J].

Kumar, CG ;

Takagi, H .

BIOTECHNOLOGY ADVANCES, 1999, 17 (07) :561-594

[9] CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[10] ANGIOTENSIN-I-CONVERTING ENZYME INHIBITORY ACTIVITIES OF SYNTHETIC PEPTIDES RELATED TO THE TANDEM REPEATED SEQUENCE OF A MAIZE ENDOSPERM PROTEIN [J].

MARUYAMA, S ;

MIYOSHI, S ;

KANEKO, T ;

TANAKA, H .

AGRICULTURAL AND BIOLOGICAL CHEMISTRY, 1989, 53 (04) :1077-1081

← 1 2 →