1887

Microbiology

Volume 143, Issue 5

Highly thermostable endo-1,3- -glucanase (laminarinase) Lam A from : nucleotide sequence of the gene and characterization of the recombinant gene product Free

Abstract

The nucleotide sequence of clone pTT26 (3786 bp), containing the gene for 1,3- -glucanase (laminarinase) from was determined. It contains an ORF encoding a protein of 646 aa (73 328 Da). The central part of the protein is homologous to the complete catalytic domain of bacterial and some eukaryotic endo-1,3- --glucanases and belongs to family 16 of glycosyl hydrolases. This domain is flanked on both sides by one copy on each side of a substrate binding domain homologue (family II). The recombinant laminarinase protein was purified from host cells in two forms, a 73 kDa and a processed 52 kDa protein, both having high specific activity towards laminarin (3100 and 2600 U mg, respectively) and values of 2.8 and 2.2 mg ml, respectively. Limited activity on 1,3-1,4- -glucan (lichenan) was detected (90 U mg). Laminarin was degraded in an endoglucanase modus, yielding glucose, laminaribiose and -triose as end products. Thus classifies as an endo-1,3(4)- -glucanase (EC 3.2.1.6). The optimum temperature of the enzymes was 95° (73 kDa) and 85° (52 kDa) at an optimum pH of 6.2. The superior thermostability of the 73 kDa enzyme is demonstrated by incubation without substrate at 100°, where 57% of the initial activity remained after 30 min (82% at 95°). Thus, is the most thermostable 1,3- -glucanase described to date.

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Keyword(s): 1,3-β-glucanase , laminarinase , thermostability and Thermotoga neapolitana
© Society Society for General Microbiology 1997
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1997-05-01
2021-10-24
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Highly thermostable endo-1,3-β -glucanase (laminarinase) Lam A from Thermotoga neapolitana: nucleotide sequence of the gene and characterization of the recombinant gene product
Microbiology 143, 1701 (1997); https://doi.org/10.1099/00221287-143-5-1701
/content/journal/micro/10.1099/00221287-143-5-1701
/content/journal/micro/10.1099/00221287-143-5-1701
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