期刊
PROTEIN AND PEPTIDE LETTERS
卷 26, 期 5, 页码 377-385出版社
BENTHAM SCIENCE PUBL LTD
DOI: 10.2174/0929866526666190228163629
关键词
Lytic polysaccharide monooxygenases; Aspergillus fumigatus; AA9 LPMO; sugarcane bagasse; biomass hydrolysis; bioethanol production
资金
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2016/19095-0]
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico [CNPq: 425465/2016-0]
- FAPESP [2018/10296-8]
- Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior - Brasil (CAPES) [001]
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)
- Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [18/10296-8, 16/19095-0] Funding Source: FAPESP
Background: Lytic Polysaccharide Monooxygenases (LPMOs) are auxiliary accessory enzymes that act synergistically with cellulases and which are increasingly being used in second-generation bioethanol production from biomasses. Several LPMOs have been identified in various filamentous fungi, including Aspergillus fumigants. However, many LPMOs have not been characterized yet. Objective: To report the role of uncharacterized A. fumigatus AfAA9_B LPMO. Methods: qRT-PCR analysis was employed to analyze the LPMO gene expression profile in different carbon sources. The gene encoding an AfAA9_B (A fu4g07850) was cloned into the vector pET-28a(+), expressed in the E. coli strain Rosetta (TM) (DE3) pLysS, and purified by a Ni2+-nitrilotriacetic (Ni-NTA) agarose resin. To evaluate the specific LPMO activity, the purified protein peroxidase activity was assessed. The auxiliary LPMO activity was investigated by the synergistic activity in Celluclast 1.5L enzymatic cocktail. Results: LPMO was highly induced in complex biomass like sugarcane bagasse (SEB), Avicel (R) PH-101, and CM-cellulose. The LPMO gene encoded a protein comprising 250 amino acids, without a CBM domain. After protein purification, the AfAA9_B molecular mass estimated by SDS-PAGE was 35 kDa. The purified protein specific peroxidase activity was 8.33+ 1.9 U g(-1). Upon addition to Celluclast 1.5L, Avicel (R) PH-101 and SEB hydrolysis increased by 18% and 22%, respectively. Conclusion: A. fumigatus LPMO is a promising candidate to enhance the currently available enzymatic cocktail and can therefore be used in second-generation ethanol production.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据