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Structural basis for the enantioselectivity of esterase Est-Y29 toward (S)-ketoprofen

Structural basis for the enantioselectivity of esterase Est-Y29 toward (S)-ketoprofen
Structural basis for the enantioselectivity of esterase Est-Y29 toward (S)-ketoprofen
The thermostable esterase Est-Y29, belonging to the family VIII lipolytic esterases isolated from metagenomes extracted from the topsoil in Republic of Korea, was identified as a promising catalyst for the production of (S)-ketoprofen, an important nonsteroidal anti-inflammatory drug (NSAID). For industrial applications, the enantioselectivity of the enzyme toward the S-enantiomer of the racemic ketoprofen ester substrate needs to be improved. To understand the structural basis of Est-Y29 enantioselectivity, which is necessary for the rational design of an enzyme with enhanced enantioselectivity, we solved the crystal structures of Est-Y29 bound to (S)-ketoprofen at 1.69 Å resolution. Structural analyses revealed that the S-enantiomer can be stabilized by a π-interaction between the methyl substituent at the chiral carbon of the ligand and the aromatic pocket formed by Tyr123, Phe125, and Tyr170. This finding is further supported by the highly improved enantioselectivity of the mutant Est-Y29 (F125W) toward (S)-ketoprofen due to the enhanced π-interaction. Our results provide the molecular basis of the enantioselectivity of Est-Y29 against (S)-ketoprofen and further offer the opportunity for the rational design of enzyme enantioselectivity as well as potential applications of the mutant Est-Y29 to industrial biocatalysts.
2155-5435
755-767
Ngo, Tri Duc
a8fc1977-0a4e-4782-a406-eb3df10f5f6a
Oh, Changsuk
34f4d821-fbf8-43c5-8753-bd29224341f3
Mizar, Pushpak
631b688c-b7fb-41d7-8038-9fb48ef935c7
Baek, Minkyung
628700ba-0bd2-4799-a821-372d85652150
Park, Kwang-Su
b6045a50-252d-42ab-b547-f2e468f3aa13
Nguyen, Lan
52413520-170c-4525-8a5e-925d4bb7195c
Byeon, Huimyoung
fac668b1-00fe-48ee-b4c4-d0cf51ba4157
Yoon, Sangyoung
324294e9-621b-4986-b4ae-2234cbae10bf
Ryu, Yeonwoo
7919180e-9d84-4c33-ad8b-0523256acaef
Ryu, Bum Han
92b453e5-2960-4750-ac98-6a5f1528773e
Kim, T. Doohun
0c1e6882-e222-4bf8-9bce-9c18a625c106
Yang, Jung Woon
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Seok, Chaok
f512d94b-0fb6-4e93-80aa-b825cc2cd7d1
Lee, Seung Seo
ee34fa26-5fb6-48c8-80c2-1f13ec4ccceb
Kim, Kyeong Kyu
a7757cb7-065e-4e76-a669-27d2108bef97
Ngo, Tri Duc
a8fc1977-0a4e-4782-a406-eb3df10f5f6a
Oh, Changsuk
34f4d821-fbf8-43c5-8753-bd29224341f3
Mizar, Pushpak
631b688c-b7fb-41d7-8038-9fb48ef935c7
Baek, Minkyung
628700ba-0bd2-4799-a821-372d85652150
Park, Kwang-Su
b6045a50-252d-42ab-b547-f2e468f3aa13
Nguyen, Lan
52413520-170c-4525-8a5e-925d4bb7195c
Byeon, Huimyoung
fac668b1-00fe-48ee-b4c4-d0cf51ba4157
Yoon, Sangyoung
324294e9-621b-4986-b4ae-2234cbae10bf
Ryu, Yeonwoo
7919180e-9d84-4c33-ad8b-0523256acaef
Ryu, Bum Han
92b453e5-2960-4750-ac98-6a5f1528773e
Kim, T. Doohun
0c1e6882-e222-4bf8-9bce-9c18a625c106
Yang, Jung Woon
70327194-b21c-402c-83c0-4474fb47db63
Seok, Chaok
f512d94b-0fb6-4e93-80aa-b825cc2cd7d1
Lee, Seung Seo
ee34fa26-5fb6-48c8-80c2-1f13ec4ccceb
Kim, Kyeong Kyu
a7757cb7-065e-4e76-a669-27d2108bef97

Ngo, Tri Duc, Oh, Changsuk, Mizar, Pushpak, Baek, Minkyung, Park, Kwang-Su, Nguyen, Lan, Byeon, Huimyoung, Yoon, Sangyoung, Ryu, Yeonwoo, Ryu, Bum Han, Kim, T. Doohun, Yang, Jung Woon, Seok, Chaok, Lee, Seung Seo and Kim, Kyeong Kyu (2019) Structural basis for the enantioselectivity of esterase Est-Y29 toward (S)-ketoprofen. ACS Catalysis, 9 (1), 755-767. (doi:10.1021/acscatal.8b02797).

Record type: Article

Abstract

The thermostable esterase Est-Y29, belonging to the family VIII lipolytic esterases isolated from metagenomes extracted from the topsoil in Republic of Korea, was identified as a promising catalyst for the production of (S)-ketoprofen, an important nonsteroidal anti-inflammatory drug (NSAID). For industrial applications, the enantioselectivity of the enzyme toward the S-enantiomer of the racemic ketoprofen ester substrate needs to be improved. To understand the structural basis of Est-Y29 enantioselectivity, which is necessary for the rational design of an enzyme with enhanced enantioselectivity, we solved the crystal structures of Est-Y29 bound to (S)-ketoprofen at 1.69 Å resolution. Structural analyses revealed that the S-enantiomer can be stabilized by a π-interaction between the methyl substituent at the chiral carbon of the ligand and the aromatic pocket formed by Tyr123, Phe125, and Tyr170. This finding is further supported by the highly improved enantioselectivity of the mutant Est-Y29 (F125W) toward (S)-ketoprofen due to the enhanced π-interaction. Our results provide the molecular basis of the enantioselectivity of Est-Y29 against (S)-ketoprofen and further offer the opportunity for the rational design of enzyme enantioselectivity as well as potential applications of the mutant Est-Y29 to industrial biocatalysts.

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Rev-sub-v2 - Accepted Manuscript
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Accepted/In Press date: 6 December 2018
e-pub ahead of print date: 24 December 2018
Published date: 1 January 2019

Identifiers

Local EPrints ID: 427176
URI: https://eprints.soton.ac.uk/id/eprint/427176
ISSN: 2155-5435
PURE UUID: 0c871e88-8072-44e1-b0e9-b960e3b06924
ORCID for Seung Seo Lee: ORCID iD orcid.org/0000-0002-8598-3303

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Date deposited: 04 Jan 2019 17:30
Last modified: 04 Apr 2019 00:31

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Contributors

Author: Tri Duc Ngo
Author: Changsuk Oh
Author: Pushpak Mizar
Author: Minkyung Baek
Author: Kwang-Su Park
Author: Lan Nguyen
Author: Huimyoung Byeon
Author: Sangyoung Yoon
Author: Yeonwoo Ryu
Author: Bum Han Ryu
Author: T. Doohun Kim
Author: Jung Woon Yang
Author: Chaok Seok
Author: Seung Seo Lee ORCID iD
Author: Kyeong Kyu Kim

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