Thermostable endoglucanase gene derived by amplification from the genomic DNA of a cellulose-enriched mixed culture from mudspring water of Mt. Makiling, Laguna, Philippines

Issue Date

3-2020

Abstract

Culture-independent molecular-based approaches can be used to identify genes of interest from environmental sources that have desirable properties such as thermo activity. For this study, a putative thermo stable endoglucanase gene was identified from a mixed culture resulting from the inoculation of Brock-CMcellulose (1%) broth with mudspring water from Mt. Makiling, Laguna, Philippines that had been incubated at 90 °C. Genomic DNA was extracted from the cellulose-enriched mixed culture and endo1949 forward and reverse primers were used to amplify the endoglucanase gene, which was cloned into pCR-script plasmid vector. Blastn alignment of the sequenced insert revealed 99.69% similarity to the glycosyl hydrolase, sso1354 (CelA1; Q97YG7) from Saccharolobus solfataricus. The endoglucanase gene (GenBank accession number MK984682) was determined to be 1,021 nucleotide bases in length, corresponding to 333 amino acids with a molecular mass of ~ 37 kDa. The endoglucanase gene was inserted into a pET21 vector and transformed in E. coli BL21 for expression. Partially purified recombinant Mt. Makiling endoglucanase (MM-Engl) showed a specific activity of 187.61 U/mg and demonstrated heat stability up to 80 °C. The thermo-acid stable endoglucanase can be used in a supplementary hydrolysis step to further hydrolyze the lignocellulosic materials that were previously treated under high temperature-dilute acid conditions, thereby enhancing the release of more glucose sugars for bioethanol production.

Source or Periodical Title

World Journal of Microbiology and Biotechnology

ISSN

0959-3993

Volume

36

Issue

51

Page

1-14

Document Type

Article

Physical Description

illustrations, graphs, references

Language

English

Subject

Endoglucanase, Lignocellulose hydrolysis, Mt. Makiling mudspring, Saccharolobus, Thermophiles

Identifier

10.1007/s11274-020-02825-2.

Digital Copy

yes

En – AGROVOC descriptors

ENDOGLUCANASE; LIGNOCELLULOSE HYDROLYSIS; MT. MAKILING MUDSPRING; SACCHAROLOBUS; THERMOPHILES

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