Purification and characterisation of acetolactate decarboxylase from Leuconostoc lactis NCW1

Siobhán M. O'Sullivan; Seamus Condon; Timothy M. Cogan; David Sheehan

doi:10.1016/S0378-1097(00)00499-7

Purification and characterisation of acetolactate decarboxylase from Leuconostoc lactis NCW1

Siobhán M. O'Sullivan, Seamus Condon, Timothy M. Cogan, David Sheehan

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11 Scopus citations

Abstract

A two-step strategy involving DEAE-cellulose and POROS PI anion exchange chromatography has been developed for rapid purification of acetolactate decarboxylase (ALD) from Leuconostoc lactis NCW1. This results in 5333-fold purification with a yield of 30%. Purified ALD is a dimer of 49-kDa subunits, has a pH optimum of 6.0, a pI of 4.2 and its activity is independent of metals or branched chain amino acids. At the optimum pH, the K_m for 2-acetolactate (ALA) was found to be 1.3 mM and the turnover number was 4000 min^-1. N-terminal sequence comparison with other ALDs showed little sequence conservation in this region. Purified ALD does not catalyse direct production of diacetyl from ALA, unlike the crude extract.

Original language	British English
Pages (from-to)	245-249
Number of pages	5
Journal	FEMS Microbiology Letters
Volume	194
Issue number	2
DOIs	https://doi.org/10.1016/S0378-1097(00)00499-7
State	Published - 15 Jan 2001

Keywords

Acetaldehyde
Acetoin
Cheese
Decarboxylase
Diacetyl
Enzyme purification
Leuconostoc

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10.1016/S0378-1097(00)00499-7

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title = "Purification and characterisation of acetolactate decarboxylase from Leuconostoc lactis NCW1",

abstract = "A two-step strategy involving DEAE-cellulose and POROS PI anion exchange chromatography has been developed for rapid purification of acetolactate decarboxylase (ALD) from Leuconostoc lactis NCW1. This results in 5333-fold purification with a yield of 30%. Purified ALD is a dimer of 49-kDa subunits, has a pH optimum of 6.0, a pI of 4.2 and its activity is independent of metals or branched chain amino acids. At the optimum pH, the Km for 2-acetolactate (ALA) was found to be 1.3 mM and the turnover number was 4000 min-1. N-terminal sequence comparison with other ALDs showed little sequence conservation in this region. Purified ALD does not catalyse direct production of diacetyl from ALA, unlike the crude extract.",

keywords = "Acetaldehyde, Acetoin, Cheese, Decarboxylase, Diacetyl, Enzyme purification, Leuconostoc",

author = "O'Sullivan, {Siobh{\'a}n M.} and Seamus Condon and Cogan, {Timothy M.} and David Sheehan",

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AU - O'Sullivan, Siobhán M.

AU - Condon, Seamus

AU - Cogan, Timothy M.

AU - Sheehan, David

PY - 2001/1/15

Y1 - 2001/1/15

N2 - A two-step strategy involving DEAE-cellulose and POROS PI anion exchange chromatography has been developed for rapid purification of acetolactate decarboxylase (ALD) from Leuconostoc lactis NCW1. This results in 5333-fold purification with a yield of 30%. Purified ALD is a dimer of 49-kDa subunits, has a pH optimum of 6.0, a pI of 4.2 and its activity is independent of metals or branched chain amino acids. At the optimum pH, the Km for 2-acetolactate (ALA) was found to be 1.3 mM and the turnover number was 4000 min-1. N-terminal sequence comparison with other ALDs showed little sequence conservation in this region. Purified ALD does not catalyse direct production of diacetyl from ALA, unlike the crude extract.

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KW - Acetaldehyde

KW - Acetoin

KW - Cheese

KW - Decarboxylase

KW - Diacetyl

KW - Enzyme purification

KW - Leuconostoc

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U2 - 10.1016/S0378-1097(00)00499-7

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SN - 0378-1097

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IS - 2

ER -

Purification and characterisation of acetolactate decarboxylase from Leuconostoc lactis NCW1

Abstract

Keywords

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