High-level expression and characterization of a novel cutinase from Malbranchea cinnamomea suitable for butyl butyrate production.
High-level expression and characterization of a novel cutinase from Malbranchea cinnamomea suitable for butyl butyrate production.
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适用于丁酸丁酯生产的肉桂霉新型角质酶的高水平表达和表征
DOI：
10.1186/s13068-017-0912-z
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发表时间：
2017
影响因子：
6.3
通讯作者：
Yang S
中科院分区：
文献类型：
--
作者：
Duan X;Liu Y;You X;Jiang Z;Yang S;Yang S
背景：丁酸丁酯是一种可由可再生和可持续的木质纤维素生物质转化的天然酯，被认为是一种很有前途的燃料来源。与传统的化学方法相比，酶法生产丁酸丁酯具有反应条件温和、专一性强、能耗低、环境友好等优点，是目前丁酸丁酯生产的最佳方法。角质酶在丁酸丁酯生产过程中起着重要作用。然而，角质酶的生产水平仍然相对较低。结果：从嗜热真菌Malbranchea cinnamomea中克隆了一个新的角质酶基因（McCut），并在毕赤酵母中进行了表达。在5L发酵罐中获得了最高酶活12，536 U/mL，是目前为止最高的角质酶产量。McCut在pH 8.0和45 °C下具有最佳活性。它在3.0-10.5的pH范围内和高达75 °C下表现出优异的稳定性。角质酶具有广泛的底物特异性，对丁酸对硝基苯基酯和三丁酸甘油酯的活性最高。它能够水解角质、聚己内酯和聚丁二酸丁二醇酯。此外，McCut还能高效合成丁酸丁酯，酯化率在4 h时达到96.9%。McCut的整体结构解析为典型的α/β-水解酶折叠。McCut与米曲霉切割酶在沟和环结构上的差异为McCut的再设计提供了有价值的信息。这些优良的特性使其在生物合成和生物降解领域具有重要的应用价值。首次对肉桂进行了鉴定和定性。高效表达P. pastorisis是迄今为止最高的角质酶。该酶具有良好的稳定性和较高的酯化效率，可用于生物燃料和化工生产。
Background:Butyl butyrate has been considered as a promising fuel source because it is a kind of natural ester which can be converted from renewable and sustainable lignocellulosic biomass. Compared with the conventional chemical methods for butyl butyrate production, the enzymatic approach has been demonstrated to be more attractive, mainly owing to the mild reaction conditions, high specificity, low energy consumption, and environmental friendliness. Cutinases play an important role in the butyl butyrate production process. However, the production level of cutinases is still relatively low. Thus, to identify novel cutinases suitable for butyl butyrate synthesis and enhance their yields is of great value in biofuel industry.Results:A novel cutinase gene (McCut) was cloned from a thermophilic fungusMalbranchea cinnamomeaand expressed inPichia pastoris. The highest cutinase activity of 12, 536 U/mL was achieved in 5-L fermentor, which is by far the highest production for a cutinase. McCut was optimally active at pH 8.0 and 45 °C. It exhibited excellent stability within the pH range of 3.0-10.5 and up to 75 °C. The cutinase displayed broad substrate specificity with the highest activity towardsp-nitrophenyl butyrate and tributyrin. It was capable of hydrolyzing cutin, polycaprolactone, and poly(butylene succinate). Moreover, McCut efficiently synthesized butyl butyrate with a maximum esterification efficiency of 96.9% at 4 h. The overall structure of McCut was resolved as a typical α/β-hydrolase fold. The structural differences between McCut andAspergillus oryzaecutinase in groove and loop provide valuable information for redesign of McCut. These excellent features make it useful in biosynthesis and biodegradation fields.Conclusions:A novel cutinase fromM. cinnamomeawas identified and characterized for the first time. High-level expression byP. pastorisis by far the highest for a cutinase. The enzyme exhibited excellent stability and high esterification efficiency for butyl butyrate production, which may make it a good candidate in biofuel and chemical industries.
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影响因子：
5
作者：
Koschorreck, Katja;Liu, Danni;Hauer, Bernhard
通讯作者：
Hauer, Bernhard
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
4.1
作者：
Calado, CRC;Ferreira, BS;Fonseca, LP
通讯作者：
Fonseca, LP
影响因子：
4
作者：
De Barros, Dragana P. C.;Azevedo, Ana M.;Fonseca, Luis P.
通讯作者：
Fonseca, Luis P.
DOI：
10.1016/j.ijbiomac.2014.07.025
发表时间：
2014-09-01
影响因子：
8.2
作者：
Fan, Guangsen;Yang, Shaoqing;Jiang, Zhengqiang
通讯作者：
Jiang, Zhengqiang