Systematic assessment of the stability of benzaldehyde lyase in aqueous-organic biphasic systems and its stabilization by modification with methoxy-poly(ethylene) glycol

被引：21

作者：

van den Wittenboer, Anne ^{[1
]}

Niemeijer, Bernd ^{[1
]}

Karmee, Sanjib Kumar ^{[2
]}

Ansorge-Schumacher, Marion B. ^{[1
,3
]}

机构：

[1] Rhein Westfal TH Aachen, Inst Biotechnol, D-52074 Aachen, Germany

[2] Rhein Westfal TH Aachen, Inst Tech & Macromol Chem, D-52074 Aachen, Germany

[3] Tech Univ Berlin, Inst Chem, Dept Enzyme Technol, D-10623 Berlin, Germany

来源：

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC | 2010年 / 67卷 / 3-4期

关键词：

Benzaldehyde lyase; Organic solvent stability; Process stability; mPEG modification; ALPHA-CHYMOTRYPSIN; 2-HYDROXY KETONES; MODIFIED ENZYMES; 2-PHASE SYSTEMS; SOLVENTS; BIOCATALYSIS; DENATURATION; ALDEHYDES; BENZOIN; LIPASE;

D O I：

10.1016/j.molcatb.2010.08.007

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：

Benzaldehyde lyase from Pseudomonas fluorescens Biovar I [BAL; E.C.4.1.2.38] catalyzes the stereoselective formation of C-C bonds coupling aldehydes to generate alpha-hydroxy ketones. A broad range of poorly water-soluble substrates are accepted in forward and reverse reactions. In this study, the stability of BAL in aqueous-organic biphasic systems as promising reaction media was systematically investigated using methyl-tert-butylether, 2-octanone, and toluene as the organic phase. Surprisingly, a strong individual molecular toxicity of these water-immiscible solvents was observed along with the interfacial toxicity exerted by the aqueous-organic interfaces. They could be considerably reduced by covalent attachment of methoxy-poly(ethylene) glycol (mPEG(750) and mPEG(2000)) to the enzyme surface increasing the half-life by a factor of up to 18. However, under reactive conditions solvent effects were strongly superimposed by an additional deactivating effect, possibly caused by the aldehyde substrate, and no differences between unmodified and modified BAL were detectable. For technical application of the enzyme in aqueous-organic biphasic media additional strategies for stabilization will therefore be desirable. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：208 / 213

页数：6

共 49 条

[1]

Ansorge-Schumacher MB, 2007, MINI-REV ORG CHEM, V4, P243

[2]

Ansorge-Schumacher Marion B., 2006, Biotechnology Journal, V1, P564, DOI 10.1002/biot.200600030

[3]

Biselli M., 2002, ENZYME CATALYSIS ORG

[4] Asymmetric synthesis of aliphatic 2-hydroxy ketones by enzymatic carboligation of aldehydes [J].

de Maria, Pablo Dominguez ;

Pohl, Martina ;

Gocke, Doerte ;

Groeger, Harald ;

Trauthwein, Harald ;

Stillger, Thomas ;

Walter, Lydia ;

Mueller, Michael .

EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, 2007, 2007 (18) :2940-2944

[5] Preparative enantio selective synthesis of benzoins and (R)-2-hydroxy-1-phenylpropanone using benzaldehyde lyase [J].

de María, PD ;

Stillger, T ;

Pohl, M ;

Wallert, S ;

Drauz, K ;

Gröger, H ;

Trauthwein, H ;

Liese, A .

JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC, 2006, 38 (01) :43-47

[6] Benzaldehyde lyase-catalyzed enantioselective carboligation of aromatic aldehydes with mono- and dimethoxy acetaldehyde [J].

Demir, AS ;

Sesenoglu, Ö ;

Dunkelmann, P ;

Muller, M .

ORGANIC LETTERS, 2003, 5 (12) :2047-2050

[7]

Demir AS, 2002, ADV SYNTH CATAL, V344, P96, DOI 10.1002/1615-4169(200201)344:1<96::AID-ADSC96>3.0.CO

[8]

2-Z

[9] Simple chemoenzymatic access to enantiopure pharmacologically interesting (R)-2-hydroxypropiophenones [J].

Demir, AS ;

Hamamci, H ;

Sesenoglu, O ;

Aydogan, F ;

Capanoglu, D ;

Neslihanoglu, R .

TETRAHEDRON-ASYMMETRY, 2001, 12 (13) :1953-1956

[10] Chemical modification of enzymes for enhanced functionality [J].

DeSantis, G ;

Jones, JB .

CURRENT OPINION IN BIOTECHNOLOGY, 1999, 10 (04) :324-330

← 1 2 3 4 5 →