Adaptation and transcriptome analysis of Aureobasidium pullulans in corncob hydrolysate for increased inhibitor tolerance to malic acid production.
Adaptation and transcriptome analysis of Aureobasidium pullulans in corncob hydrolysate for increased inhibitor tolerance to malic acid production.
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DOI：
10.1371/journal.pone.0121416
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Wu X
中科院分区：
文献类型：
--
作者：
Zou X;Wang Y;Tu G;Zan Z;Wu X
苹果酸是一种广泛应用于食品工业的二元酸，也是一种潜在的C4平台化学品。玉米芯是一种低成本的农业可再生原料。然而，在玉米芯前处理过程中会产生严重阻碍发酵的副反应产物(呋喃、5-羟甲基呋喃、甲酸和醋酸)。以一株产多聚马来酸(PMA)的出芽短梗霉为研究对象，研究了玉米芯水解液生产苹果酸的工艺。在最佳水解液糖浓度为110g/L的条件下，通过逐步提高水解液中的糖浓度，使出芽孢杆菌在好氧纤维床生物反应器中得到进一步的适应。经过9批发酵，苹果酸的产量和产量分别达到38.6g/L和0.4g/L h，高于第一批的27.6g/L和0.29g/L h。适应菌株能在0.5g/L糠醛、3g/L羟甲基纤维素钠、2g/L冰醋酸、0.5g/L甲酸的胁迫下生长，而野生菌株则不能生长。转录组分析表明，在0.5g/L和3g/L两种胁迫条件下，差异表达基因分别与糖类运输和代谢、脂类运输和代谢、信号转导机制、氧化还原代谢以及能量产生和转化相关。在适应株中，42个基因的表达水平上调了15倍以上，qRT-PCR也证实了与硫同化途径有关的关键基因(即SIR、GSS、CyS和GSR)的表达水平在适应株中上调了10倍以上。
Malic acid is a dicarboxylic acid widely used in the food industry, and is also a potential C4 platform chemical. Corncob is a low-cost renewable feedstock from agricultural industry. However, side-reaction products (furfural, 5-hydroxymethylfurfural (HMF), formic acid, and acetic acid) that severely hinder fermentation are formed during corncob pretreatment. The process for producing malic acid from a hydrolysate of corncob was investigated with a polymalic acid (PMA)-producing Aureobasidium pullulans strain. Under the optimal hydrolysate sugar concentration 110 g/L, A. pullulans was further adapted in an aerobic fibrous bed bioreactor (AFBB) by gradually increasing the sugar concentration of hydrolysate. After nine batches of fermentation, the production and productivity of malic acid reached 38.6 g/L and 0.4 g/L h, respectively, which was higher than that in the first batch (27.6 g/L and 0.29 g/L h, respectively). The adapted strain could grow under the stress of 0.5 g/L furfural, 3 g/L HMF, 2g/L acetic acid, and 0.5 g/L formic acid, whereas the wild type did not. Transcriptome analysis revealed that the differentially expressed genes were related to carbohydrate transport and metabolism, lipid transport and metabolism, signal transduction mechanism, redox metabolism, and energy production and conversion under 0.5 g/L furfural and 3 g/L HMF stress conditions. In total, 42 genes in the adapted strain were upregulated by 15-fold or more, and qRT-PCR also confirmed that the expression levels of key genes (i.e. SIR, GSS, CYS, and GSR) involved in sulfur assimilation pathway were upregulated by over 10-fold in adapted strain for cellular protection against oxidative stress.
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