Wu Na, Xing Mingyan, Chen Yaru, Zhang Chi, Li Yingfeng, Song Ping, Xu Qing, Liu Hao, Huang He
School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210023, China.
College of Life Sciences, Nanjing Normal University, Nanjing, 210046, China.
Biotechnol Biofuels Bioprod. 2022 Dec 29;15(1):151. doi: 10.1186/s13068-022-02250-7.
As an attractive platform chemical, malic acid has been commonly used in the food, feed and pharmaceutical field. Microbial fermentation of biobased sources to produce malic acid has attracted great attention because it is sustainable and environment-friendly. However, most studies mainly focus on improving yield and ignore shortening fermentation time. A long fermentation period means high cost, and hinders the industrial applications of microbial fermentation. Stresses, especially oxidative stress generated during fermentation, inhibit microbial growth and production, and prolong fermentation period. Previous studies have shown that polypeptides could effectively relieve stresses, but the underlying mechanisms were poorly understood.
In this study, polypeptides (especially elastin peptide) addition improves the productivity of malic acid in A. niger, resulting in shortening of fermentation time from 120 to 108 h. Transcriptome and biochemical analyses demonstrated that both antioxidant enzyme-mediated oxidative stress defense system, such as superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX), and nonenzymatic antioxidant system, such as glutathione, were enhanced in the presence of elastin peptide, suggesting elastin peptide relieving oxidative stresses is involved in many pathways. In order to further investigate the relationship between oxidative stress defense and malic acid productivity, we overexpressed three enzymes (Sod1, CAT, Tps1) related to oxidation resistance in A. niger, respectively, and these resulting strains display varying degree of improvement in malic acid productivity. Especially, the strain overexpressing the Sod1 gene achieved a malate titer of 91.85 ± 2.58 g/L in 96 h, corresponding to a productivity of 0.96 g/L/h, which performs better than elastin peptide addition.
Our investigation provides an excellent reference for alleviating the stress of the fungal fermentation process and improving fermentation efficiency.
苹果酸作为一种具有吸引力的平台化学品,已广泛应用于食品、饲料和制药领域。利用生物基原料通过微生物发酵生产苹果酸因其可持续性和环境友好性而备受关注。然而,大多数研究主要集中在提高产量上,而忽视了缩短发酵时间。较长的发酵周期意味着高成本,并阻碍了微生物发酵的工业应用。压力,尤其是发酵过程中产生的氧化应激,会抑制微生物的生长和产物合成,并延长发酵周期。先前的研究表明,多肽可以有效缓解压力,但其潜在机制尚不清楚。
在本研究中,添加多肽(尤其是弹性蛋白肽)提高了黑曲霉中苹果酸的生产效率,使发酵时间从120小时缩短至108小时。转录组和生化分析表明,在弹性蛋白肽存在的情况下,抗氧化酶介导的氧化应激防御系统,如超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPX),以及非酶抗氧化系统,如谷胱甘肽,均得到增强,这表明弹性蛋白肽缓解氧化应激涉及多种途径。为了进一步研究氧化应激防御与苹果酸生产效率之间的关系,我们分别在黑曲霉中过表达了三种与抗氧化相关的酶(Sod1、CAT、Tps1),这些产生的菌株在苹果酸生产效率上有不同程度的提高。特别是,过表达Sod1基因的菌株在96小时内实现了苹果酸滴度为91.85±2.58 g/L,对应生产效率为0.96 g/L/h,表现优于添加弹性蛋白肽的情况。
我们的研究为缓解真菌发酵过程中的压力和提高发酵效率提供了很好的参考。