Transcriptome analysis of Aureobasidium pullulans BL06 and identification of key factors affecting pullulan production

IF 10.7 1区 化学 Q1 CHEMISTRY, APPLIED Carbohydrate Polymers Pub Date : 2024-11-13 DOI:10.1016/j.carbpol.2024.122984
Jiaqi Gao , Xiaoping Liao , Hongwu Ma , Wenqin Bai
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Abstract

Pullulan, a versatile water-soluble polysaccharide, is widely used across various industries. To minimize byproduct interference, Aureobasidium pullulans BL06ΔPMAs was engineered, resulting in a higher yield and a lower molecular weight (MW) than the parent strain A. pullulans BL06. Comparative transcriptomic analysis revealed differentially expressed genes (DEGs) involved in sucrose metabolism, gluconeogenesis, glyoxylate metabolism, and amino acid metabolism. These DEGs may influence substrate consumption, energy production, and membrane composition, ultimately impacting pullulan synthesis. Additionally, further experimental validations were conducted on the genes with the most significant differential expression. Overexpressing glycosyltransferase gene (gta1, the third most differentially expressed gene) in A. pullulans BL06 increased pullulan production by 8.1 %, indicating its role in short α-1,4-glucan synthesis. Overexpression of the transmembrane transporter gene (st1, the most significantly differentially expressed gene) reduced pullulan molecular weight by 25 %, which potentially influences cell membrane fluidity and pullulan secretion. Furthermore, amylase (Amy1) was found to significantly impact molecular weight (MW) within the first 48 h of fermentation, an effect not previously reported for amylase, while its knockout resulted in a remarkable 7.6-fold increase in pullulan MW. These findings provide valuable insights for regulating pullulan yield and MW, offering innovative genetic targets for strains engineering in future industrial applications.
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Aureobasidium pullulans BL06 的转录组分析以及影响乌拉坦生产的关键因素的鉴定
普鲁兰是一种多功能水溶性多糖,广泛应用于各行各业。为了最大限度地减少副产品干扰,对 Aureobasidium pullulans BL06ΔPMAs 进行了工程化改造,从而获得了比亲本菌株 A. pullulans BL06 更高的产量和更低的分子量(MW)。比较转录组分析揭示了涉及蔗糖代谢、葡萄糖生成、乙醛酸代谢和氨基酸代谢的差异表达基因(DEGs)。这些 DEGs 可能会影响底物消耗、能量产生和膜组成,并最终影响拉伸聚糖的合成。此外,还对差异表达最显著的基因进行了进一步的实验验证。在 A. pullulans BL06 中,过表达糖基转移酶基因(gta1,差异表达第三大的基因)可使拉胶产量增加 8.1%,表明其在短α-1,4-葡聚糖合成中的作用。过量表达跨膜转运体基因(st1,差异表达最显著的基因)会使拉胶分子量降低 25%,这可能会影响细胞膜的流动性和拉胶的分泌。此外,研究还发现淀粉酶(Amy1)在发酵的最初 48 小时内对分子量(MW)有显著影响,这是以前未报道过的淀粉酶的影响,而淀粉酶基因敲除则使毛胶分子量显著增加了 7.6 倍。这些发现为调节拉毛丹产量和分子量提供了宝贵的见解,为未来工业应用中的菌株工程提供了创新的遗传目标。
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来源期刊
Carbohydrate Polymers
Carbohydrate Polymers 化学-高分子科学
CiteScore
22.40
自引率
8.00%
发文量
1286
审稿时长
47 days
期刊介绍: Carbohydrate Polymers stands as a prominent journal in the glycoscience field, dedicated to exploring and harnessing the potential of polysaccharides with applications spanning bioenergy, bioplastics, biomaterials, biorefining, chemistry, drug delivery, food, health, nanotechnology, packaging, paper, pharmaceuticals, medicine, oil recovery, textiles, tissue engineering, wood, and various aspects of glycoscience. The journal emphasizes the central role of well-characterized carbohydrate polymers, highlighting their significance as the primary focus rather than a peripheral topic. Each paper must prominently feature at least one named carbohydrate polymer, evident in both citation and title, with a commitment to innovative research that advances scientific knowledge.
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