Gene cloning, expression and biochemical characterization of a glucose- and xylose-stimulated β-glucosidase from Humicola insolens RP86

纤维二糖 木糖 化学 水解 生物化学 纤维素酶 β-葡萄糖苷酶 重组DNA 发酵 基因
作者
Flávio Henrique Moreira Souza,Luana Parras Meleiro,Carla Botelho Machado,Ana Lúcia Ribeiro Latorre Zimbardi,Raquel Fonseca Maldonado,T.A.C.B. Souza,Douglas Chodi Masui,M.T. Murakami,João Atı́lio Jorge,Richard J. Ward,Rosa dos Prazeres Melo Furriel
出处
期刊:Journal of Molecular Catalysis B-enzymatic [Elsevier BV]
卷期号:106: 1-10 被引量:33
标识
DOI:10.1016/j.molcatb.2014.04.007
摘要

Efficient cellulose saccharification is crucial for the cost-effective production of ethanol from lignocellulosic biomass. Most β-glucosidases are inhibited by glucose, and high enzyme loads are needed to attain acceptable hydrolysis yields, increasing the process costs. Therefore, glucose-stimulated β-glucosidases with high catalytic efficiency for cellobiose hydrolysis are highly interesting, particularly those produced at low cost using convenient expression systems. The bglhi gene from Humicola insolens RP86, which encodes a glucose- and xylose-stimulated β-glucosidase (Bglhi), was cloned and expressed in E. coli. The recombinant Bglhi was expressed in soluble form and dynamic light scattering showed that the protein was monomeric (56.1 kDa). The enzyme showed broad substrate specificity, with optima of pH and temperature of 5.0–7.0 and 60 °C, and was stable from pH 4.5 to 8.0, and for 1 h at 50 °C in water. The recombinant Bglhi was highly stimulated by glucose and xylose. About 2-fold stimulation of pNP-glucosidase activity occurred at 50 mmol L−1 glucose or 100 mmol L−1 xylose. In addition, stimulation was observed over 50–370 mmol L−1 glucose or 100–1000 mmol L−1 xylose. Xylose maximally stimulated (2.3-fold) the cellobiase activity of the enzyme at 80 mmol L−1 concentration. The Vmax for cellobiose hydrolysis (183.4 ± 12.0 U mg−1) was 5-fold higher than that for pNP-Glc (36.4 ± 1.4 U mg−1), but a lower KM for pNP-Glc (0.20 ± 0.01 mmol L−1), compared to cellobiose (0.38 ± 0.01 mmol L−1), was estimated. More importantly, the recombinant Bglhi presented the highest catalytic efficiency for cellobiose hydrolysis (kcat/KM = 453.0 ± 29.6 s−1 mmol−1 L) among all glucose-stimulated and glucose-tolerant β-glucosidases known to date. Together, these properties indicate that the recombinant Bglhi is an attractive candidate to include in enzymatic cocktails for application in industrial processes for the saccharification of lignocellulosic materials.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
聪明的宛菡完成签到,获得积分10
1秒前
CNYDNZB完成签到 ,获得积分10
1秒前
xxj完成签到 ,获得积分10
1秒前
芊芊完成签到 ,获得积分10
2秒前
yar应助bluesky采纳,获得10
2秒前
海人完成签到 ,获得积分10
3秒前
SY15732023811完成签到 ,获得积分10
5秒前
李建勋完成签到,获得积分10
5秒前
科研通AI2S应助一路芬芳采纳,获得10
5秒前
黄花完成签到 ,获得积分10
6秒前
刘珍荣完成签到,获得积分10
7秒前
7秒前
紫金之巅完成签到 ,获得积分10
7秒前
Gang完成签到,获得积分10
8秒前
9秒前
9秒前
10秒前
CYYDNDB完成签到 ,获得积分10
10秒前
粿粿一定行完成签到 ,获得积分10
11秒前
12秒前
战战完成签到,获得积分10
13秒前
xlk2222完成签到,获得积分10
16秒前
笨笨以莲完成签到,获得积分10
16秒前
YHX完成签到,获得积分10
17秒前
沐沐心完成签到 ,获得积分10
18秒前
18秒前
19秒前
哭泣笑柳发布了新的文献求助10
19秒前
轻松白桃完成签到,获得积分10
19秒前
JasVe完成签到 ,获得积分10
22秒前
wakkkkk完成签到,获得积分10
22秒前
含蓄听南完成签到,获得积分10
22秒前
HH给HH的求助进行了留言
22秒前
芋你呀完成签到,获得积分10
23秒前
西蓝花香菜完成签到 ,获得积分10
23秒前
无花果应助兔子采纳,获得10
23秒前
请勿继续完成签到,获得积分10
25秒前
搞怪的婴完成签到,获得积分10
26秒前
Loooong完成签到,获得积分0
26秒前
28秒前
高分求助中
【提示信息,请勿应助】关于scihub 10000
Les Mantodea de Guyane: Insecta, Polyneoptera [The Mantids of French Guiana] 3000
徐淮辽南地区新元古代叠层石及生物地层 3000
The Mother of All Tableaux: Order, Equivalence, and Geometry in the Large-scale Structure of Optimality Theory 3000
Handbook of Industrial Diamonds.Vol2 1100
Global Eyelash Assessment scale (GEA) 1000
Picture Books with Same-sex Parented Families: Unintentional Censorship 550
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 4038184
求助须知:如何正确求助?哪些是违规求助? 3575908
关于积分的说明 11373872
捐赠科研通 3305715
什么是DOI,文献DOI怎么找? 1819255
邀请新用户注册赠送积分活动 892662
科研通“疑难数据库(出版商)”最低求助积分说明 815022