Characterization of β-glucosidases from Meridianimaribacter sp. CL38: Characterization of β-glucosidases from Meridianimaribacter sp. CL38

Clarine Wan Ling Hong; Sye Jinn Chen; Kok Jun Liew; Ming Quan Lam; Muhammad Ramziuddin Zakaria; Kheng Loong Chong; Chun Shiong Chong

doi:10.11594/jtls.13.03.09

Authors

Clarine Wan Ling Hong Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, Skudai 81310, Malaysia
Sye Jinn Chen Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, Skudai 81310, Malaysia
Kok Jun Liew Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, Skudai 81310, Malaysia
Ming Quan Lam Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, Skudai 81310, Malaysia
Muhammad Ramziuddin Zakaria Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, Skudai 81310, Malaysia
Kheng Loong Chong Department of Biosciences, Faculty of Science, Universiti Teknologi Malaysia, Skudai 81310, Malaysia
Chun Shiong Chong RM 108, Level 3, Block T02, Faculty of Science (FS), Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Malaysia.

DOI:

https://doi.org/10.11594/jtls.13.03.09

Keywords:

β-glucosidase, GH3, Halophiles, Lignocellulosic biomass, Meridianimaribacter

Abstract

The production of second-generation biofuel requires a huge amount of freshwater. It is estimated that at least three gal of freshwater is used to produce one gal of biofuel. The replacement of freshwater with seawater serves as a potential alternative in biofuel generation. Therefore, salt-tolerant enzymes play an important role in saccharification and fermentation process. Halophilic β-glucosidase is one of the key enzymes for the process. In this study, the β-glucosidase of halophile Meridianimaribacter sp. CL38 isolated from mangrove soil was characterized. Strain CL38 achieved maximum production of β-glucosidase at 12^th hour of growth. The β-glucosidase showed highest activity at 2% (w/v) NaCl while highly stable at salt concentration ranging from 1-2% (w/v) (more than 96% of relative activity). Its β-glucosidase activity remained active in the presence of 5mM Mn²⁺, Mg²⁺, Ca²⁺ ions, and 1% (v/v) Tween-20 and Tween-80. The draft genome sequence of strain CL38 was retrieved from GenBank database and submitted to dbCAN meta server for CAZymes annotation. Strain CL38 harbors 44 GHs and GH3 are annotated as β-glucosidases. The β-glucosidases of Meridianimaribacter flavus (99.61%) and Mesoflavibacter sabulilitoris (97.44%) showed the closest identity with Bgl3a and Bgl3b protein sequences from strain CL38, respectively. Glycoside hydrolase family 3 domain was identified in both enzymes via InterPro scan server. The presence of signal peptides indicated that both enzymes were secreted extracellularly. Five motifs were identified in Bgl3a and Bgl3b, with the active site (nucleophile) found at Asp296 and Asp297, respectively. Collectively, these β-glucosidases could be potentially used in the biofuel production, in particular the lignocellulosic biomass pretreatment process. This is the first attempt to characterize the β-glucosidase in genus Meridianimaribacter as so far none of the lignocellulolytic enzymes from this genus were characterized.

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