Production of propane and other short-chain alkanes by structure-based engineering of ligand specificity in aldehyde-deformylating oxygenase.
Production of propane and other short-chain alkanes by structure-based engineering of ligand specificity in aldehyde-deformylating oxygenase.
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DOI：
10.1002/cbic.201300307
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发表时间：
2013-07-08
期刊：
影响因子：
3.2
通讯作者：
Scrutton, Nigel S.
中科院分区：
文献类型：
--
作者：
Khara, Basile;Menon, Navya;Levy, Colin;Mansell, David;Das, Debasis;Marsh, E. Neil G.;Leys, David;Scrutton, Nigel S.
近年来，蓝藻已成为有吸引力的微生物宿主烃生产。[1,2]然而，在蓝藻细菌中生产生物燃料的可行性仍然具有挑战性，需要识别和工程的天然生物催化剂来生产烷烃并将其整合到代谢过程中。[b蓝藻的碳氢化合物生物合成被认为是由脂肪酸分解代谢引起的，涉及两个主要的酶促反应：酰基载体蛋白还原酶（AAR，将脂肪酸转化为醛），然后羰基的丧失形成碱(e)，由醛脱甲酰氧合酶（ADO）催化，以前称为醛脱羰酶（AD）；方案1)[4-7]已经分离出蓝藻ADOs (cADOs)，并显示其可以脱碳长链醛（C18及以上），这是假定的生理底物，同时也显示出与中链醛（如庚醛）的体外活性。[8,9]人们对改造蓝藻以积累短链烷烃并产生“滴入”燃料（如丙烷）有着浓厚的兴趣。这就需要酶催化剂能够从中枢代谢产生的前体中产生这些碳氢化合物。在这里，我们着手通过基于结构的基质通道工程来设计海洋原胆球菌（菌株MIT9313） cADO的改进变体。我们的目的是改变cADO的天然特异性，使其对短链醛的反应性高于对长链醛的反应性，从而为代谢工程生产新的催化模块。
In recent years, cyanobacteria have emerged as attractive microbial hosts for hydrocarbon production.[1, 2] However, whilst appealing, the practicalities of producing biofuels in cyanobacteria remain challenging, requiring the identification and engineering of natural biocatalysts for alkane production and their integration into metabolic processes.[3] Cyanobacterial hydrocarbon biosynthesis is presumed to arise from fatty acid catabolism involving two main enzymatic reactions, an acyl carrier protein reductase (AAR; converting fatty acids to aldehydes) followed by loss of the carbonyl group to form alka (e) nes catalysed by an aldehyde-deformylating oxygenase (ADO), known formerly as aldehyde decarbonylase (AD; Scheme 1).[4–7] Cyanobacterial ADOs (cADOs) have been isolated and shown to decarbonylate long-chain aldehydes (C18 and above), which are the presumed physiological substrates, whilst also displaying in vitro activity with medium-chain aldehydes (eg, heptanal).[8, 9]There is intense interest in engineering cyanobacteria to accumulate short-chain alkanes and produce “drop in” fuels such as propane. This requires enzyme catalysts that can produce these hydrocarbons from precursors derived from central metabolism. Here, we set out to engineer improved variants of Procholorococcus marinus (strain MIT9313) cADO through structure-based engineering of the substrate-access tunnel. Our intention was to alter the natural specificity of cADO to favour reactivity against short-chain over long-chain aldehydes, and thereby produce new catalytic modules for metabolic engineering.
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影响因子：
2.9
作者：
Li, Ning;Chang, Wei-chen;Bollinger, J. Martin, Jr.
通讯作者：
Bollinger, J. Martin, Jr.
通过非血红素铁依赖性蓝藻醛脱羰酶校正不依赖于氧的烷烃形成：动力学研究和外部电子供体的需求。
影响因子：
2.9
作者：
Eser BE;Das D;Han J;Jones PR;Marsh EN
通讯作者：
Marsh EN
影响因子：
7.8
作者：
Krebs, Carsten;Bollinger, J. Martin, Jr.;Booker, Squire J.
通讯作者：
Booker, Squire J.
影响因子：
15
作者：
Warui, Douglas M.;Li, Ning;Norgaard, Hanne;Krebs, Carsten;Bollinger, J. Martin, Jr.;Booker, Squire J.
通讯作者：
Booker, Squire J.
DOI：
10.1073/pnas.89.12.5306
发表时间：
1992-06-15
影响因子：
11.1
作者：
DENNIS, M;KOLATTUKUDY, PE
通讯作者：
KOLATTUKUDY, PE