Cloning of Poly(aspartic acid) (PAA) Hydrolase-1 Gene from Pedobacter sp KP-2 and Hydrolysis of Thermally Synthesized PAA by its Gene Product

被引：12

作者：

Hiraishi, Tomohiro ^{[1
]}

Masuda, Eriko ^{[1
,2
]}

Kanayama, Naoki ^{[1
]}

Nagata, Madoka ^{[3
]}

Doi, Yoshiharu ^{[3
]}

Abe, Hideki ^{[2
,3
]}

Maeda, Mizuo ^{[1
]}

机构：

[1] Riken Inst Phys & Chem Res, Bioengn Lab, Wako, Saitama 3510198, Japan

[2] Tokyo Inst Technol, Dept Innovat & Engn Mat, Midori Ku, Kanagawa 2268502, Japan

[3] Riken Inst Phys & Chem Res, Polymer Chem Lab, Wako, Saitama 3510198, Japan

来源：

MACROMOLECULAR BIOSCIENCE | 2009年 / 9卷 / 01期

关键词：

beta-aspartic acid; biodegradable; pedobacter sp KP-2; poly(aspartic acid) hydrolase; water-soluble polymers; SPHINGOMONAS SP KT-1; L-ASPARTIC ACID; ESCHERICHIA COLI; POLY(SUCCINIMIDE); BIODEGRADABILITY; POLYCONDENSATION; POLYASPARTATE; PURIFICATION; DEGRADATION; PHOSPHATASE;

D O I：

10.1002/mabi.200800106

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：

Pedobacter sp. KP-2 can degrade and metabolize thermally synthesized alpha,beta-poly(D,L-aspartic acid) (tPAA), which contains 70% of unnatural beta-amide units, with high-molecular-weight. In this study, gene cloning and molecular characterization of PAA hydrolase-1 from KP-2 was carried out. Gene analysis reveals that deduced amino acid sequence of the enzyme shows a similarity to only that of PAA hydrolase-1 from Sphingomonas sp. KT-1. GPC and NMR analyses of the hydrolyzed products of tPAA by PAA hydrolase-1 of KP-2 indicate that this enzyme cleaves the beta-beta amide linkage via endo-mode to yield oligo(aspartic acid) from tPAA. Taking the composition of tPAA and the substrate specificity of PAA hydrolase-1 into consideration, the enzyme possibly plays a crucial role in tPAA biodegradation by KP-2.

引用

页码：10 / 19

页数：10

共 30 条

[1] THE LOCATION OF DISSIMILATORY NITRITE REDUCTASE AND THE CONTROL OF DISSIMILATORY NITRATE REDUCTASE BY OXYGEN IN PARACOCCUS-DENITRIFICANS [J].

ALEFOUNDER, PR ;

FERGUSON, SJ .

BIOCHEMICAL JOURNAL, 1980, 192 (01) :231-240

[2]

Alford Diana D., 1994, Journal of Environmental Polymer Degradation, V2, P225, DOI 10.1007/BF02071970

[3]

[Anonymous], 1989, Molecular Cloning: A Laboratory Manual

[4]

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

[5] ON LOCALIZATION OF ALKALINE PHOSPHATASE AND CYCLIC PHOSPHODIESTERASE IN ESCHERICHIA COLI [J].

BROCKMAN, RW ;

HEPPEL, LA .

BIOCHEMISTRY, 1968, 7 (07) :2554-+

[6] PURIFICATION AND PROPERTIES OF 2 ACID PHOSPHATASE FRACTIONS ISOLATED FROM OSMOTIC SHOCK FLUID OF ESCHERICHIA COLI [J].

DVORAK, HF ;

BROCKMAN, RW ;

HEPPEL, LA .

BIOCHEMISTRY, 1967, 6 (06) :1743-+

[7]

Freeman MB, 1996, ACS SYM SER, V627, P118

[8] Enzymatic hydrolysis of α- and β-oligo(L-aspartic acid)s by poly(aspartic acid) hydrolases-1 and 2 from Sphingromonas sp KT-1 [J].

Hiraishi, T ;

Kajiyama, M ;

Yamato, K ;

Doi, Y .

MACROMOLECULAR BIOSCIENCE, 2004, 4 (03) :330-339

[9] Biochemical and molecular characterization of poly(aspartic acid) hydrolase-2 from Sphingomonas sp KT-1 [J].

Hiraishi, T ;

Kajiyama, M ;

Tabata, K ;

Abe, H ;

Yamato, I ;

Doi, Y .

BIOMACROMOLECULES, 2003, 4 (05) :1285-1292

[10] Genetic analysis and characterization of poly(aspartic acid) hydrolase-1 from Sphingomonas sp KT-1 [J].

Hiraishi, T ;

Kajiyama, M ;

Tabata, K ;

Yamato, I ;

Doi, Y .

BIOMACROMOLECULES, 2003, 4 (01) :80-86

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