Implications of cellobiohydrolase glycosylation for use in biomass conversion.
Implications of cellobiohydrolase glycosylation for use in biomass conversion.
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细胞胞胞成虫酶糖基化对生物量转化的影响。
DOI：
10.1186/1754-6834-1-10
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发表时间：
2008-05-01
影响因子：
6.3
通讯作者：
Adney, William S.
中科院分区：
文献类型：
--
作者：
Jeoh, Tina;Michener, William;Himmel, Michael E.;Decker, Stephen R.;Adney, William S.
已知产生纤维素酶的子囊菌里氏木霉（Hypocrea jecorina）能够分泌一系列对于从木质纤维素生物质生产乙醇很重要的酶。由于其能够生产高滴度的异源蛋白质，它也被广泛用于工业酶的商业规模生产。然而，在分泌过程中，可能会发生许多影响蛋白质功能和稳定性的翻译后事件。另一种子囊菌，黑曲霉变种。泡盛还可以为工业生产大量异源蛋白质。在这项研究中，里氏木霉 Cel7A（一种纤维二糖水解酶）在黑曲霉变种中表达。泡盛并进行详细的生物物理表征。纯化的重组酶含有的 N-连接聚糖数量是从商业里氏木霉酶制剂中纯化的酶的六倍。使用细菌（微晶）和磷酸溶胀（无定形）纤维素作为底物比较两种酶形式的活性。这一比较表明，重组 Cel7A (rCel7A) 的 N-糖基化水平增加导致活性降低和对纤维素的非生产性结合增加。当用N-糖苷酶PNGaseF处理时，重组酶的分子量接近商业酶的分子量，并且对纤维素的活性得到提高。
The cellulase producing ascomycete, Trichoderma reesei (Hypocrea jecorina), is known to secrete a range of enzymes important for ethanol production from lignocellulosic biomass. It is also widely used for the commercial scale production of industrial enzymes because of its ability to produce high titers of heterologous proteins. During the secretion process, a number of post-translational events can occur, however, that impact protein function and stability. Another ascomycete, Aspergillus niger var. awamori, is also known to produce large quantities of heterologous proteins for industry. In this study, T. reesei Cel7A, a cellobiohydrolase, was expressed in A. niger var. awamori and subjected to detailed biophysical characterization. The purified recombinant enzyme contains six times the amount of N-linked glycan than the enzyme purified from a commercial T. reesei enzyme preparation. The activities of the two enzyme forms were compared using bacterial (microcrystalline) and phosphoric acid swollen (amorphous) cellulose as substrates. This comparison suggested that the increased level of N-glycosylation of the recombinant Cel7A (rCel7A) resulted in reduced activity and increased non-productive binding on cellulose. When treated with the N-glycosidase PNGaseF, the molecular weight of the recombinant enzyme approached that of the commercial enzyme and the activity on cellulose was improved.
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影响因子：
6.2
作者：
Helbert, W;Chanzy, H;Ernst, S
通讯作者：
Ernst, S
影响因子：
3.4
作者：
Krystynowicz, A;Czaja, W;Bielecki, S
通讯作者：
Bielecki, S
影响因子：
4.1
作者：
NUMMI, M;NIKUPAAVOLA, ML;RAUNIO, V
通讯作者：
RAUNIO, V
影响因子：
8.4
作者：
Den Haan, Riaan;Rose, Shaunita H.;van Zyl, Willem H.
通讯作者：
van Zyl, Willem H.
DOI：
10.1038/nbt0885-722
发表时间：
1985-01-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
HENRISSAT, B;DRIGUEZ, H;SCHULEIN, M
通讯作者：
SCHULEIN, M