Raw starch conversion by Saccharomyces cerevisiae expressing Aspergillus tubingensis amylases.

Raw starch conversion by Saccharomyces cerevisiae expressing Aspergillus tubingensis amylases.
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DOI:
10.1186/1754-6834-6-167
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发表时间:
2013-11-29
影响因子:
6.3
通讯作者:
Viljoen-Bloom M
Viljoen-Bloom M
中科院分区:
工程技术1区
文献类型:
--
作者:
Viktor MJ;Rose SH;van Zyl WH;Viljoen-Bloom M

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淀粉是最丰富的有机多糖之一,可用于生产生物乙醇作为替代运输燃料。淀粉的成本效益利用需要综合生物加工(CBP),其中单个微生物可以产生淀粉水解所需的酶,并将葡萄糖单体转化为乙醇。塔宾曲霉 T8.4 α-淀粉酶 (amyA) 和葡糖淀粉酶 (glaA) 基因在实验室菌株酿酒酵母 Y294 和半工业菌株酿酒酵母 Mnuα1 中克隆并表达。重组 AmyA 和 GlaA 的蛋白质大小分别为 110-150 kDa 和 90 kDa,表明酿酒酵母中存在显着的糖基化。 10 天后,Mnuα1[AmyA-GlaA] 和 Y294[AmyA-GlaA] 菌株能够利用 20 g l-1 生玉米淀粉作为唯一碳水化合物来源,乙醇滴度分别为 9.03 和 6.67 g l-1(0.038 和 0.028 g l-1 h-1)。在底物负载为 200 g l-1 生玉米淀粉的情况下,Mnuα1[AmyA-GlaA] 在发酵 120 h 后产生 70.07 g l-1 乙醇 (0.58 g l-1 h-1),而 Y294[AmyA-GlaA] 的效率较低,产生 43.33 g l-1 乙醇(0.36 g l-1 h-1)。在表达 Tubingensis α-淀粉酶和葡糖淀粉酶基因的半工业化淀粉分解酿酒酵母菌株中,200 g l-1 生淀粉在 120 小时内完全水解(糖化),其中 74% 转化为释放的糖和发酵产物,其余部分可能转化为生物质。生淀粉的一步转化代表了在无需任何热预处理的情况下实现 CBP 的重大进展。此外,淀粉酶是由宿主菌株产生和分泌的,从而避免了对外源淀粉酶的需要。
Starch is one of the most abundant organic polysaccharides available for the production of bio-ethanol as an alternative transport fuel. Cost-effective utilisation of starch requires consolidated bioprocessing (CBP) where a single microorganism can produce the enzymes required for hydrolysis of starch, and also convert the glucose monomers to ethanol. The Aspergillus tubingensis T8.4 α-amylase (amyA) and glucoamylase (glaA) genes were cloned and expressed in the laboratory strain Saccharomyces cerevisiae Y294 and the semi-industrial strain, S. cerevisiae Mnuα1. The recombinant AmyA and GlaA displayed protein sizes of 110–150 kDa and 90 kDa, respectively, suggesting significant glycosylation in S. cerevisiae. The Mnuα1[AmyA-GlaA] and Y294[AmyA-GlaA] strains were able to utilise 20 g l-1 raw corn starch as sole carbohydrate source, with ethanol titers of 9.03 and 6.67 g l-1 (0.038 and 0.028 g l-1 h-1), respectively, after 10 days. With a substrate load of 200 g l-1 raw corn starch, Mnuα1[AmyA-GlaA] yielded 70.07 g l-1 ethanol (0.58 g l-1 h-1) after 120 h of fermentation, whereas Y294[AmyA-GlaA] was less efficient at 43.33 g l-1 ethanol (0.36 g l-1 h-1). In a semi-industrial amylolytic S. cerevisiae strain expressing the A. tubingensis α-amylase and glucoamylase genes, 200 g l-1 raw starch was completely hydrolysed (saccharified) in 120 hours with 74% converted to released sugars plus fermentation products and the remainder presumably to biomass. The single-step conversion of raw starch represents significant progress towards the realisation of CBP without the need for any heat pretreatment. Furthermore, the amylases were produced and secreted by the host strain, thus circumventing the need for exogenous amylases.
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发表时间: 2010-03-01
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影响因子: 2.3
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DOI: 10.1016/0922-338x(96)80579-4
发表时间: 1996-01-01
期刊: JOURNAL OF FERMENTATION AND BIOENGINEERING
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