Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter.
Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter.
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DOI：
10.1186/1754-6834-7-8
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发表时间：
2014-01-14
影响因子：
6.3
通讯作者：
Kondo A
中科院分区：
文献类型：
--
作者：
Inokuma K;Hasunuma T;Kondo A
利用糖基磷脂酰肌醇(GPI)锚定系统在细胞表面展示异源纤维素酶的重组酵母菌株被认为是木质纤维原料直接转化为乙醇的很有前途的生物催化剂。然而，传统的纤维素酶酵母菌株对纤维素酶的分解活性不足以降解纤维素酶。在本研究中，我们通过展示纤维素酶的酵母菌株构建了高效利用纤维素的新型基因盒。利用酿酒酵母SED1GPI锚定区和启动子构建了尖锐曲霉β-葡萄糖苷酶(BGL1)和里氏木霉内切葡聚糖酶II(EGII)的细胞面展示基因盒。将基因盒整合到酿酒酵母基因组中，对重组菌株的β-葡萄糖苷酶活性进行了检测。我们发现，同时利用基因盒中的SED1启动子和Sed1锚定结构域能够高效地将酶整合到细胞壁中。转导新基因盒的重组酵母细胞的β-葡萄糖苷酶活性是常规菌株的8.4倍。展示EGII的新型菌株对水不溶纤维素的水解活性也是常规菌株的106倍。此外，通过使用新的基因盒展示里氏木霉EGII基因，提高了水热处理稻草直接生产乙醇的能力。我们开发了新的基因盒，用于高效地展示外型和内型纤维素酶的细胞表面。结果表明，该基因盒具有广泛的细胞表面展示的适用性，纤维素酶展示酵母在利用木质纤维生物质生产生物乙醇方面具有巨大的潜力。
The recombinant yeast strains displaying the heterologous cellulolytic enzymes on the cell surface using the glycosylphosphatidylinositol (GPI) anchoring system are considered promising biocatalysts for direct conversion of lignocellulosic materials to ethanol. However, the cellulolytic activities of the conventional cellulase-displaying yeast strains are insufficient for the hydrolysis of cellulose. In this study, we constructed novel gene cassettes for the efficient cellulose utilization by cellulase-displaying yeast strains. The novel gene cassettes for the cell-surface display of Aspergillus aculeatus β-glucosidase (BGL1) and Trichoderma reeseii endoglucanase II (EGII) were constructed using the promoter and the GPI anchoring region derived from Saccharomyces cerevisiae SED1. The gene cassettes were integrated into the S. cerevisiae genome, then the β-glucosidase activity of these recombinant strains was evaluated. We revealed that simultaneous utilization of the SED1 promoter and Sed1 anchoring domain in a gene cassette enabled highly-efficient enzyme integration into the cell wall. The β-glucosidase activity of recombinant yeast cells transduced with the novel gene cassette was 8.4-fold higher than that of a conventional strain. The novel EGII-displaying strain also achieved 106-fold higher hydrolysis activity against the water-insoluble cellulose than a conventional strain. Furthermore, direct ethanol production from hydrothermally processed rice straw was improved by the display of T. reeseii EGII using the novel gene cassette. We have developed novel gene cassettes for the efficient cell-surface display of exo- and endo-type cellulolytic enzymes. The results suggest that this gene cassette has the wide applicability for cell-surface display and that cellulase-displaying yeasts have significant potential for cost-effective bioethanol production from lignocellulosic biomass.
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影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
影响因子：
5
作者：
Katahira, Satoshi;Mizuike, Atsuko;Kondo, Akihiko
通讯作者：
Kondo, Akihiko
影响因子：
3.5
作者：
Cha-aim, Kamonchai;Fukunaga, Tomoaki;Akada, Rinji
通讯作者：
Akada, Rinji
影响因子：
6.3
作者：
Lau MW;Gunawan C;Balan V;Dale BE
通讯作者：
Dale BE
影响因子：
11.4
作者：
Doering, TL;Schekman, R
通讯作者：
Schekman, R