Production of squalene in Synechocystis sp. PCC 6803.
Production of squalene in Synechocystis sp. PCC 6803.
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DOI：
10.1371/journal.pone.0090270
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Lindberg P
中科院分区：
文献类型：
--
作者：
Englund E;Pattanaik B;Ubhayasekera SJ;Stensjö K;Bergquist J;Lindberg P
近年来，人们对研究和开发化石燃料的可持续替代品越来越感兴趣。使用光合微生物来生产这样的替代品是有利的，因为它们可以利用阳光作为能源，将大气中的二氧化碳直接转化为所需的产品。角鲨烯是一种天然存在的30碳类异戊二烯，在化妆品和疫苗中具有商业用途。如果可以大规模可持续地生产，它也可以代替石油作为燃料的原材料和化学工业的原料。单细胞蓝细菌集胞藻PCC 6803具有预测编码角鲨烯环化酶（Shc）的基因slr 2089，所述角鲨烯环化酶是将角鲨烯转化成环化烯的酶，环化烯是形成类霍帕烷的底物。通过对slr2089（shc）的失活，探索利用蓝藻生产角鲨烯的可能性。失活导致角鲨烯的积累，其水平比野生型细胞高70倍以上，达到0.67 mg OD 750 −1 L−1。与野生型集胞藻相比，我们没有观察到任何显著的生长缺陷，即使在高光条件下，这表明观察到的角鲨烯积累对生长无害，并且Shc形成的hopene对于正常条件下的生长和高光胁迫耐受性都不是至关重要的。研究了不同光照强度和生长阶段对Δshc菌株角鲨烯积累的影响。我们还确定了一个基因，sll0513，作为一个假定的角鲨烯合酶在集胞藻，并验证了其功能的失活。在这项工作中，我们表明，这是可能的，使用蓝藻集胞藻产生角鲨烯，碳氢化合物的商业利益和潜在的生物燃料。我们还报告了第一次鉴定的角鲨烯环化酶，和第二次鉴定的角鲨烯合酶，在蓝藻。
In recent years, there has been an increased interest in the research and development of sustainable alternatives to fossil fuels. Using photosynthetic microorganisms to produce such alternatives is advantageous, since they can achieve direct conversion of carbon dioxide from the atmosphere into the desired product, using sunlight as the energy source. Squalene is a naturally occurring 30-carbon isoprenoid, which has commercial use in cosmetics and in vaccines. If it could be produced sustainably on a large scale, it could also be used instead of petroleum as a raw material for fuels and as feedstock for the chemical industry. The unicellular cyanobacterium Synechocystis PCC 6803 possesses a gene, slr2089, predicted to encode squalene hopene cyclase (Shc), an enzyme converting squalene into hopene, the substrate for forming hopanoids. Through inactivation of slr2089 (shc), we explored the possibility to produce squalene using cyanobacteria. The inactivation led to accumulation of squalene, to a level over 70 times higher than in wild type cells, reaching 0.67 mg OD750 −1 L−1. We did not observe any significant growth deficiency in the Δshc strain compared to the wild type Synechocystis, even at high light conditions, suggesting that the observed squalene accumulation was not detrimental to growth, and that formation of hopene by Shc is not crucial for growth under normal conditions, nor for high-light stress tolerance. Effects of different light intensities and growth stages on squalene accumulation in the Δshc strain were investigated. We also identified a gene, sll0513, as a putative squalene synthase in Synechocystis, and verified its function by inactivation. In this work, we show that it is possible to use the cyanobacterium Synechocystis to generate squalene, a hydrocarbon of commercial interest and a potential biofuel. We also report the first identification of a squalene hopene cyclase, and the second identification of squalene synthase, in cyanobacteria.
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影响因子：
3.9
作者：
Lichtenthaler, HK
通讯作者：
Lichtenthaler, HK
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
影响因子：
4.8
作者：
Gu, PD;Ishii, Y;Shechter, I
通讯作者：
Shechter, I
影响因子：
3.2
作者：
Barkley, SJ;Desai, SB;Poulter, CD
通讯作者：
Poulter, CD
DOI：
10.1111/j.1432-1033.1996.0051r.x
发表时间：
1996-11-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Feil, C;Sussmuth, R;Poralla, K
通讯作者：
Poralla, K