Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine

The lipase B from Candida antarctica (CalB)-catalyzed enantioselective resolution of N-(2-ethyl-6-methylphenyl)alanine (NEMPA) from the corresponding racemic methyl ester has been investigated. However, the enantioselectivity of CalB towards the resolution is not high enough to obtain enantiomerical...

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Main Authors: Zheng, Liangyu, Zhang, Suoqin, Han, Yu
Format: Text
Language:unknown
Published: ECI Digital Archives 2017
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Online Access:https://dc.engconfintl.org/enzyme_xxiv/40
id fteci:oai:dc.engconfintl.org:enzyme_xxiv-1139
record_format openpolar
spelling fteci:oai:dc.engconfintl.org:enzyme_xxiv-1139 2023-05-15T14:04:06+02:00 Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine Zheng, Liangyu Zhang, Suoqin Han, Yu 2017-09-24T07:00:00Z https://dc.engconfintl.org/enzyme_xxiv/40 unknown ECI Digital Archives https://dc.engconfintl.org/enzyme_xxiv/40 Enzyme Engineering XXIV Candida antarctica lipase B Kinetic resolution NEMPA Rational regulation Enantioselectivity Engineering text 2017 fteci 2022-12-27T14:49:04Z The lipase B from Candida antarctica (CalB)-catalyzed enantioselective resolution of N-(2-ethyl-6-methylphenyl)alanine (NEMPA) from the corresponding racemic methyl ester has been investigated. However, the enantioselectivity of CalB towards the resolution is not high enough to obtain enantiomerically pure products. With the aim to improve lipase CalB enantioselectivity, the effects of microenvironment on lipase CalB-catalyzed enantioselective resolution were initially investigated, and the high enantioselective ratio of CalB (e.e.p=90.5%) is achieved in THF/buffer (35% v/v). Introduce additives such as amides and amino acids in THF/water reaction system, the enantioselective ratio of CalB can be improved further (e.e.p is up to 98.6%). Structural studies with IR spectroscopy and Michaelis–Menten kinetics show that the established strong hydrogen bonds interactions between enzyme and solvents are the main driving force that affects the spatial conformation of the enzyme, which further affect the affinity and enantioselectivity of lipase CalB towards the substrate. Based on these results, we modified the lipase CalB with amine-rich molecules to increase the hydrogen interactions between lipase CalB, reaction solvents and substrates. The resulted lipase CalB tagged with 6-histidine at the N terminal and 10-lysine at the C terminal (6His-CalB-10Lys) indeed show the better enantioselectivity than that of commercial lipase CalB in enantioselective resolution of NEMPA in pure buffer. In particular, polyamine tags fused in CalB can also help achieve high soluble expression of CalB in E. coli and can directly mediate silicification, which leads to rapid formation of a CalB-silica particle complex through a one-step approach. The strategies described are practical process for obtaining high enantiomerically pure (S)-NEMPA using lipase CalB. This abstract is an original research focused on the enhancement of the enantioselectivity of lipase CalB-catalyzed hydrolysis reaction by rational change of solvents, addition of ... Text Antarc* Antarctica Engineering Conferences International: ECI Digital Archives
institution Open Polar
collection Engineering Conferences International: ECI Digital Archives
op_collection_id fteci
language unknown
topic Candida antarctica lipase B
Kinetic resolution
NEMPA
Rational regulation
Enantioselectivity
Engineering
spellingShingle Candida antarctica lipase B
Kinetic resolution
NEMPA
Rational regulation
Enantioselectivity
Engineering
Zheng, Liangyu
Zhang, Suoqin
Han, Yu
Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine
topic_facet Candida antarctica lipase B
Kinetic resolution
NEMPA
Rational regulation
Enantioselectivity
Engineering
description The lipase B from Candida antarctica (CalB)-catalyzed enantioselective resolution of N-(2-ethyl-6-methylphenyl)alanine (NEMPA) from the corresponding racemic methyl ester has been investigated. However, the enantioselectivity of CalB towards the resolution is not high enough to obtain enantiomerically pure products. With the aim to improve lipase CalB enantioselectivity, the effects of microenvironment on lipase CalB-catalyzed enantioselective resolution were initially investigated, and the high enantioselective ratio of CalB (e.e.p=90.5%) is achieved in THF/buffer (35% v/v). Introduce additives such as amides and amino acids in THF/water reaction system, the enantioselective ratio of CalB can be improved further (e.e.p is up to 98.6%). Structural studies with IR spectroscopy and Michaelis–Menten kinetics show that the established strong hydrogen bonds interactions between enzyme and solvents are the main driving force that affects the spatial conformation of the enzyme, which further affect the affinity and enantioselectivity of lipase CalB towards the substrate. Based on these results, we modified the lipase CalB with amine-rich molecules to increase the hydrogen interactions between lipase CalB, reaction solvents and substrates. The resulted lipase CalB tagged with 6-histidine at the N terminal and 10-lysine at the C terminal (6His-CalB-10Lys) indeed show the better enantioselectivity than that of commercial lipase CalB in enantioselective resolution of NEMPA in pure buffer. In particular, polyamine tags fused in CalB can also help achieve high soluble expression of CalB in E. coli and can directly mediate silicification, which leads to rapid formation of a CalB-silica particle complex through a one-step approach. The strategies described are practical process for obtaining high enantiomerically pure (S)-NEMPA using lipase CalB. This abstract is an original research focused on the enhancement of the enantioselectivity of lipase CalB-catalyzed hydrolysis reaction by rational change of solvents, addition of ...
format Text
author Zheng, Liangyu
Zhang, Suoqin
Han, Yu
author_facet Zheng, Liangyu
Zhang, Suoqin
Han, Yu
author_sort Zheng, Liangyu
title Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine
title_short Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine
title_full Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine
title_fullStr Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine
title_full_unstemmed Rational enhancement of the enantioselectivity of Candida antarctica lipase B in kinetic resolution of N-(2-ethyl-6-methylphenyl) alanine
title_sort rational enhancement of the enantioselectivity of candida antarctica lipase b in kinetic resolution of n-(2-ethyl-6-methylphenyl) alanine
publisher ECI Digital Archives
publishDate 2017
url https://dc.engconfintl.org/enzyme_xxiv/40
genre Antarc*
Antarctica
genre_facet Antarc*
Antarctica
op_source Enzyme Engineering XXIV
op_relation https://dc.engconfintl.org/enzyme_xxiv/40
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