LIPASE OF PSEUDOMONAS-CEPACIA FOR BIOTECHNOLOGICAL PURPOSES - PURIFICATION, CRYSTALLIZATION AND CHARACTERIZATION

被引：68

作者：

BORNSCHEUER, U

REIF, OW

LAUSCH, R

FREITAG, R

SCHEPER, T

KOLISIS, FN

MENGE, U

机构：

[1] GESELL BIOTECHNOL FORSCH MBH,D-38124 BRAUNSCHWEIG,GERMANY

[2] SCH AGR SCI,DEPT APPL BIOL SCI,MOLEC BIOL SCI LAB,CHIKUSA KU,NAGOYA,AICHI 46401,JAPAN

[3] INST TECH CHEM,D-30167 HANNOVER 1,GERMANY

[4] UNIV MUNSTER,INST BIOCHEM,BIOTECHNOL ABT,D-48149 MUNSTER,GERMANY

[5] NATL TECH UNIV ATHENS,DEPT CHEM ENGN,GR-15700 ATHENS,GREECE

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 1994年 / 1201卷 / 01期

关键词：

TRIACYLGLYCEROL LIPASE; PROTEIN PURIFICATION; (P-CEPACIA);

D O I：

10.1016/0304-4165(94)90151-1

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：

Commercial lipase (triacylglycerol lipase, EC 3.1.1.3) of Pseudomonas cepacia (Amano) has been purified to homogeneity by a single chromatography on phenyl Sepharose. The eluted lipase crystallized spontaneously at 4 degrees C in the eluent, containing 58-69% 2-propanol. The yield of the lipase was 87-100% and the specific activity during the hydrolysis of triolein 5800 U/mg protein. This protein has a molecular weight of 34.1 kDa as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Its purity was determined by SDS-PAGE and capillary zone electrophoresis to be greater than or equal to 99%. Immobilization on Sepharose increased its stability in organic solvents. This lipase of P. cepacia differs from that of other Pseudomonas strains in respect to substrate specificity and during crystallization. It exhibits a high stability in organic solvents and supercritical carbon dioxide.

引用

页码：55 / 60

页数：6

共 23 条

[1] INFLUENCES OF REACTION CONDITIONS ON THE ENANTIOSELECTIVE TRANSESTERIFICATION USING PSEUDOMONAS-CEPACIA LIPASE
BORNSCHEUER, U
SCHAPOHLER, S
SCHEPER, T
SCHUGERL, K
[J]. TETRAHEDRON-ASYMMETRY, 1991, 2 (10) : 1011 - 1014
[2] BORNSCHEUER U, 1992, ANN NY ACAD SCI, V672, P336
[3] FACTORS AFFECTING THE LIPASE-CATALYZED TRANSESTERIFICATION REACTIONS OF 3-HYDROXY ESTERS IN ORGANIC-SOLVENTS
BORNSCHEUER, U
HERAR, A
KREYE, L
WENDEL, V
CAPEWELL, A
MEYER, HH
SCHEPER, T
KOLISIS, FN
[J]. TETRAHEDRON-ASYMMETRY, 1993, 4 (05) : 1007 - 1016
[4] BORNSCHEUER U, 1987, VDI17 FORTSCHR BER
[5] BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6] SUBSTRATE SPECIFICITIES OF LIPASE-A AND LIPASE-B FROM GEOTRICHUM-CANDIDUM CMICC-335426
CHARTON, E
MACRAE, AR
[J]. BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1123 (01) : 59 - 64
[7] CLONING OF THE PSEUDOMONAS-GLUMAE LIPASE GENE AND DETERMINATION OF THE ACTIVE-SITE RESIDUES
FRENKEN, LGJ
EGMOND, MR
BATENBURG, AM
BOS, JW
VISSER, C
VERRIPS, CT
[J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1992, 58 (12) : 3787 - 3791
[8] SIMPLIFIED METHOD FOR SILVER STAINING OF PROTEINS IN POLYACRYLAMIDE GELS AND THE MECHANISM OF SILVER STAINING
HEUKESHOVEN, J
DERNICK, R
[J]. ELECTROPHORESIS, 1985, 6 (03) : 103 - 112
[9] SPARSE-MATRIX SAMPLING - A SCREENING METHOD FOR CRYSTALLIZATION OF PROTEINS
JANCARIK, J
KIM, SH
[J]. JOURNAL OF APPLIED CRYSTALLOGRAPHY, 1991, 24 : 409 - 411
[10] Jensen R.G., 1974, LIPOLYTIC ENZYMES, P139

← 1 2 3 →