Improved secretory expression of lignocellulolytic enzymes in Kluyveromyces marxianus by promoter and signal sequence engineering.
Improved secretory expression of lignocellulolytic enzymes in Kluyveromyces marxianus by promoter and signal sequence engineering.
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通过启动子和信号序列工程改善马克斯克鲁维酵母中木质纤维素分解酶的分泌表达
DOI：
10.1186/s13068-018-1232-7
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发表时间：
2018
影响因子：
6.3
通讯作者：
Yu Y
中科院分区：
文献类型：
--
作者：
Zhou J;Zhu P;Hu X;Lu H;Yu Y
背景：考虑到它的耐热性、高生长速度和广泛的底物光谱，马氏克鲁维菌可以被认为是理想的巩固生物处理（CBP）。使用k生产乙醇的一个主要障碍。马氏菌的木质纤维素水解酶产量低，这降低了纤维素水解和乙醇的产量。因此，进一步改善酶的表达和分泌是必要的。结果：菊粉酶启动子和k基因的信号序列提高了木质素纤维素水解酶的表达。通过诱变对马氏菌进行了优化。AT（-361）启动子内的突变，5'UTR内AT-rich区域的缺失（UTR∆A）以及信号序列中P10L的替换使阿铁酰酯酶Est1E的分泌表达增加了6倍。T(-361)A和UTR∆A增加了mRNA的表达，而P10L的取代延长了信号序列的疏水核心，促进了成熟蛋白的分泌。P10L和T(-361)A突变使其他类型的木质纤维素水解酶的分泌表达增加了三倍，包括endo1,4 -β-葡聚糖酶RuCelA， endo1,4 -β-内生木聚糖酶Xyn-CDBFV和endo1,4 -β-甘露聚糖酶MAN330。在携带优化模块的菌株补料分批发酵过程中，RuCelA、xn - cdbfv、MAN330和Est1E的活性峰值分别达到24 U/mL、25,600 U/mL、10,200 U/mL和1220 U/mL。重要的是，优化的启动子和信号序列在所有测试的碳源中都实现了更高的酶产量，包括木质纤维素糖化和发酵的主要最终产物，在木糖上生长的酶产量最高。结论：基因工程启动子和信号序列可增加不同木质纤维素水解酶inK的分泌表达。马克思利用各种碳资源。饲料分批发酵中木质纤维素水解酶活性最高。marxianusso远。我们的工程模块在生产byK的木质纤维素水解酶方面很有价值。构建高效的纤维素乙醇生产CBP菌株。
Background:Taking into account its thermotolerance, high growth rate, and broad substrate spectrum,Kluyveromyces marxianuscan be considered an ideal consolidated bioprocessing (CBP). A major obstacle to ethanol production usingK. marxianusis the low production of lignocellulolytic enzymes, which reduces the cellulose hydrolysis and ethanol production. Thus, further improvement of enzyme expression and secretion is essential.Results:To improve the expression of lignocellulolytic enzymes, the inulinase promoter and signal sequence fromK. marxianuswas optimized through mutagenesis. A T(-361)A mutation inside the promoter, a deletion of AT-rich region inside 5'UTR (UTR∆A), and a P10L substitution in the signal sequence increased the secretory expression of the feruloyl esterase Est1E by up to sixfold. T(-361)A and UTR∆A increased the mRNA expression, while the P10L substitution extended the hydrophobic core of signal sequence and promoted secretion of mature protein. P10L and T(-361)A mutations increased secretory expressions of other types of lignocellulolytic enzymes by up to threefold, including endo-1,4-β-glucanase RuCelA, endo-1,4-β-endoxylanase Xyn-CDBFV, and endo-1,4-β-mannanase MAN330. During the fed-batch fermentation of strains carrying optimized modules, the peak activities of RuCelA, Xyn-CDBFV, MAN330, and Est1E reached 24 U/mL, 25,600 U/mL, 10,200 U/mL, and 1220 U/mL, respectively. Importantly, higher yield of enzymes by optimized promoter and signal sequence were achieved in all tested carbon sources, including the major end products of lignocellulose saccharification and fermentation, with growth on xylose resulting in the highest production.Conclusions:The engineered promoter and signal sequence presented increased secretory expressions of different lignocellulolytic enzymes inK. marxianusby means of various carbon resources. Activities of lignocellulolytic enzymes in fed-batch fermentation were the highest activities reported forK. marxianusso far. Our engineered modules are valuable in producing lignocellulolytic enzymes byK. marxianusand in constructing efficient CBP strains for cellulosic ethanol production.
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前胰岛素原信号肽的裂解受损与常染色体显性糖尿病有关
影响因子：
7.7
作者：
Liu M;Lara-Lemus R;Shan SO;Wright J;Haataja L;Barbetti F;Guo H;Larkin D;Arvan P
通讯作者：
Arvan P
DOI：
10.1073/pnas.93.11.5208
发表时间：
1996-05-28
影响因子：
11.1
作者：
Iyer, V;Struhl, K
通讯作者：
Struhl, K
影响因子：
4.1
作者：
NUMMI, M;NIKUPAAVOLA, ML;RAUNIO, V
通讯作者：
RAUNIO, V
影响因子：
5
作者：
Chang, Lei;Ding, Mozhu;Lu, Hong
通讯作者：
Lu, Hong
影响因子：
3.2
作者：
Gao, Jiaoqi;Yuan, Wenjie;Jiang, Yu
通讯作者：
Jiang, Yu