Cloning, expression, characterization and role of the leader sequence of a lipase from Rhizopus oryzae

被引：57
作者：
Beer, HD
McCarthy, JEG
Bornscheuer, UT
Schmid, RD
机构：
[1] Univ Stuttgart, Inst Tech Biochem, D-70569 Stuttgart, Germany
[2] MPI Biochem, AG Werner, D-82152 Martinsried, Germany
[3] UMIST, Dept Biochem, Manchester M60 1QD, Lancs, England
来源：
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION | 1998年 / 1399卷 / 2-3期
关键词：
lipase; leader sequence; cloning; expression; sequence; (Rhizopus oryzae);
D O I：
10.1016/S0167-4781(98)00104-3
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
A lipase from Rhizopus oryzae DSM 853 (ROL) was cloned from a chromosomal gene bank, sequenced and overexpressed in Escherichia coli. ROL and its precursors ProROL and PreProROL were purified and their pH and temperature profile was determined. In contrast to ROL, ProROL and PreProROL had considerably higher thermostability and a slightly higher pH optimum. Moreover, it could be demonstrated by in vitro experiments that the natural leader sequence of ROL is able to inhibit the folding supporting properties of the prosequence, resulting in a retardation of folding. In addition, there is strong evidence that all different lipase forms derived from Rhizopus sp. described in the literature are a result of different proteolytic processing and originate from the same gene. (C) 1998 Elsevier Science B.V. All rights reserved.
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收藏
页码：173 / 180
页数：8
相关论文
共 29 条
[1]   The role of pro regions in protein folding [J].
Baker, David ;
Shiau, Andrew K. ;
Agard, David A. .
CURRENT OPINION IN CELL BIOLOGY, 1993, 5 (06) :966-970
[2]   SEQUENCES IMPORTANT FOR GENE-EXPRESSION IN FILAMENTOUS FUNGI [J].
BALLANCE, DJ .
YEAST, 1986, 2 (04) :229-236
[3]   Analysis of the catalytic mechanism of a fungal lipase using computer-aided design and structural mutants [J].
Beer, HD ;
Wohlfahrt, G ;
McCarthy, JEG ;
Schomburg, D ;
Schmid, RD .
PROTEIN ENGINEERING, 1996, 9 (06) :507-517
[4]   RHIZOMUCOR-MIEHEI TRIGLYCERIDE LIPASE IS SYNTHESIZED AS A PRECURSOR [J].
BOEL, E ;
HUGEJENSEN, B ;
CHRISTENSEN, M ;
THIM, L ;
FIIL, NP .
LIPIDS, 1988, 23 (07) :701-706
[5]   LIPASE OF PSEUDOMONAS-CEPACIA FOR BIOTECHNOLOGICAL PURPOSES - PURIFICATION, CRYSTALLIZATION AND CHARACTERIZATION [J].
BORNSCHEUER, U ;
REIF, OW ;
LAUSCH, R ;
FREITAG, R ;
SCHEPER, T ;
KOLISIS, FN ;
MENGE, U .
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1994, 1201 (01) :55-60
[6]  
BREATHNACH R, 1981, ANNU REV BIOCHEM, V50, P349, DOI 10.1146/annurev.bi.50.070181.002025
[7]   TRIGGER FACTOR - A SOLUBLE-PROTEIN THAT FOLDS PRO-OMPA INTO A MEMBRANE-ASSEMBLY-COMPETENT FORM [J].
CROOKE, E ;
WICKNER, W .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (15) :5216-5220
[8]   PERIPLASMIC PRODUCTION OF CORRECTLY PROCESSED HUMAN GROWTH-HORMONE IN ESCHERICHIA-COLI - NATURAL AND BACTERIAL SIGNAL SEQUENCES ARE INTERCHANGEABLE [J].
GRAY, GL ;
BALDRIDGE, JS ;
MCKEOWN, KS ;
HEYNEKER, HL ;
CHANG, CN .
GENE, 1985, 39 (2-3) :247-254
[9]   CLONING, EXPRESSION AND CHARACTERIZATION OF A CDNA-ENCODING A LIPASE FROM RHIZOPUS-DELEMAR [J].
HAAS, MJ ;
ALLEN, J ;
BERKA, TR .
GENE, 1991, 109 (01) :107-113
[10]   EVOLUTIONARY CONSERVATION OF COMPONENTS OF THE PROTEIN TRANSLOCATION COMPLEX [J].
HARTMANN, E ;
SOMMER, T ;
PREHN, S ;
GORLICH, D ;
JENTSCH, S ;
RAPOPORT, TA .
NATURE, 1994, 367 (6464) :654-657
123