TY - JOUR
T1 - Characterization and high-level expression of a metagenome-derived alkaline pectate lyase in recombinant Escherichia coli
AU - Wang, Huilin
AU - Li, Xiaoman
AU - Ma, Yanhe
AU - Song, Jiangning
PY - 2014
Y1 - 2014
N2 - Alkaline pectate lyases (PLs) play an important role in mild and eco-friendly bioscouring pretreatment processes in the textile industry. However, to date, only a few PLs can be applied in industrial-scale production, and many of them exhibit high production cost, low activity, and/or do not meet the treatment requirements. In this study, an alkaline PL gene was cloned from the metagenomic DNA of alkaline environment soils. The gene pelB consisted of 1263 nucleotides and encoded a mature protein (PelB) of 399 amino acids, which was expressed in Escherichia coli. The maximum catalytic activity of the enzyme exhibited a bimodal distribution at pH 8.1 and 9.8 and an optimal temperature of 55?C. The Km and Vmax
values of PelB were 1.78 g/L and 1084.8 mol/(L min) at 45?C, respectively. Substrate speci?city analysis exhibited a bimodal distribution at pH 8.1 and 9.8 and an optimal temperature of 55?C. The Km and Vmax values of PelB were 1.78 g/L and 1084.8 mol/(L min) at 45?C, respectively. Substrate speci?city analysis cell-density cultivation in 7-L bioreactor, the highest PL activity (1816.2 U/mL) was achieved. Thus, the recombinant PelB, with promising properties for use in bioscouring in the textile pretreatment process
should be a potential enzyme for industrial applications.
AB - Alkaline pectate lyases (PLs) play an important role in mild and eco-friendly bioscouring pretreatment processes in the textile industry. However, to date, only a few PLs can be applied in industrial-scale production, and many of them exhibit high production cost, low activity, and/or do not meet the treatment requirements. In this study, an alkaline PL gene was cloned from the metagenomic DNA of alkaline environment soils. The gene pelB consisted of 1263 nucleotides and encoded a mature protein (PelB) of 399 amino acids, which was expressed in Escherichia coli. The maximum catalytic activity of the enzyme exhibited a bimodal distribution at pH 8.1 and 9.8 and an optimal temperature of 55?C. The Km and Vmax
values of PelB were 1.78 g/L and 1084.8 mol/(L min) at 45?C, respectively. Substrate speci?city analysis exhibited a bimodal distribution at pH 8.1 and 9.8 and an optimal temperature of 55?C. The Km and Vmax values of PelB were 1.78 g/L and 1084.8 mol/(L min) at 45?C, respectively. Substrate speci?city analysis cell-density cultivation in 7-L bioreactor, the highest PL activity (1816.2 U/mL) was achieved. Thus, the recombinant PelB, with promising properties for use in bioscouring in the textile pretreatment process
should be a potential enzyme for industrial applications.
UR - http://www.sciencedirect.com/science/article/pii/S135951131300562X
U2 - 10.1016/j.procbio.2013.10.001
DO - 10.1016/j.procbio.2013.10.001
M3 - Article
SN - 1359-5113
VL - 49
SP - 69
EP - 76
JO - Process Biochemistry
JF - Process Biochemistry
IS - 1
ER -