Miao Jiaxi, Wang Mengmeng, Ma Lei, Li Tuo, Huang Qiwei, Liu Dongyang, Shen Qirong
Jiangsu Key Lab for Organic Solid Waste Utilization, Nanjing, 210095 China.
Jiangsu Collaborative Innovation Center for Solid Organic Waste Resource Utilization, Nanjing, 210095 China.
Biotechnol Biofuels. 2019 Jan 4;12:4. doi: 10.1186/s13068-018-1350-2. eCollection 2019.
As a ubiquitous filamentous fungal, spp. play a critical role in lignocellulose degradation, which was also defined as considerable cell factories for organic acids and industrially relevant enzymes producer. Nevertheless, the production of various extracellular enzymes can be influenced by different factors including nitrogen source, carbon source, cultivation temperature, and initial pH value. Thus, this study aims to reveal how amino acids affect the decomposition of lignocellulose by Z5 through transcriptional and proteomics methods.
The activities of several lignocellulosic enzymes secreted by Z5 adding with cysteine, methionine, and ammonium sulfate were determined with the chromatometry method. The peak of endo-glucanase (7.33 ± 0.03 U mL), exo-glucanase (10.50 ± 0.07 U mL), β-glucosidase (21.50 ± 0.22 U mL), and xylanase (76.43 ± 0.71 U mL) were all obtained in the Cys treatment. The secretomes of Z5 under different treatments were also identified by LC-MS/MS, and 227, 256 and 159 different proteins were identified in the treatments of Cys, Met, and CK (Control, treatment with ammonium sulfate as the sole nitrogen source), respectively. Correlation analysis results of transcriptome and proteome data with fermentation profiles showed that most of the cellulose-degrading enzymes including cellulases, hemicellulases and glycoside hydrolases were highly upregulated when cysteine was added to the growth medium. In particular, the enzymes that convert cellulose into cellobiose appear to be upregulated. This study could increase knowledge of lignocellulose bioconversion pathways and fungal genetics.
Transcriptome and proteome analyses' results indicated that cysteine could significantly promote the secretion of lignocellulosic enzymes of an efficient lignocellulosic decomposing strain, Z5. The possible reason for these results is that Z5 preferred to use amino acids such as cysteine to adapt to the external environment through upregulating carbon-related metabolism pathways.
作为一种广泛存在的丝状真菌,[具体菌种名称]在木质纤维素降解中起着关键作用,其也被定义为生产有机酸和工业相关酶的重要细胞工厂。然而,各种胞外酶的产生会受到包括氮源、碳源、培养温度和初始pH值等不同因素的影响。因此,本研究旨在通过转录组学和蛋白质组学方法揭示氨基酸如何影响[菌种名称]Z5对木质纤维素的分解。
采用比色法测定了添加半胱氨酸、蛋氨酸和硫酸铵的[菌种名称]Z5分泌的几种木质纤维素酶的活性。内切葡聚糖酶(7.33±0.03 U/mL)、外切葡聚糖酶(10.50±0.07 U/mL)、β-葡萄糖苷酶(21.50±0.22 U/mL)和木聚糖酶(76.43±0.71 U/mL)的活性峰值均在半胱氨酸处理中获得。还通过液相色谱-串联质谱法(LC-MS/MS)鉴定了不同处理下[菌种名称]Z5的分泌蛋白组,在半胱氨酸、蛋氨酸和CK(对照,以硫酸铵作为唯一氮源的处理)处理中分别鉴定出227、256和159种不同的蛋白质。转录组和蛋白质组数据与发酵谱的相关性分析结果表明,当向生长培养基中添加半胱氨酸时,包括纤维素酶、半纤维素酶和糖苷水解酶在内的大多数纤维素降解酶均高度上调。特别是,将纤维素转化为纤维二糖的酶似乎被上调。本研究可以增加对木质纤维素生物转化途径和真菌遗传学的认识。
转录组和蛋白质组分析结果表明,半胱氨酸可以显著促进高效木质纤维素分解菌株[菌种名称]Z5的木质纤维素酶分泌。这些结果的可能原因是[菌种名称]Z5倾向于使用半胱氨酸等氨基酸,通过上调碳相关代谢途径来适应外部环境。