Recombinant β-agarases: insights into molecular, biochemical, and physiochemical characteristics

Abstract

Agarases (agarose 4-glycanohydrolase; EC 3.2.1.81) are class of enzymes that belong to glycoside hydrolase (GH) family capable of hydrolyzing agar. Their classification depends on hydrolysis pattern and product formation. Among all the agarases, β-agarases and the oligosaccharides formed by its action have fascinated quite a lot of industries. Ample of β-agarase genes have been endowed from marine sources such as algae, sea water, and marine sediments, and the expression of these genes into suitable host gives rise to recombinant β-agarases. These recombinant β-agarases have wide range of industrial applications due to its improved catalytic efficiency and stability in tough environments with ease of production on large scale. In this review, we have perused different types of recombinant β-agarases in consort with their molecular, physiochemical, and kinetic properties in detail and the significant features of those agarases are spotlighted. From the literature reviewed after 2010, we have found that the recombinant β-agarases belonged to the families GH16, GH39, GH50, GH86, and GH118. Among that, GH39, GH50, and GH86 belonged to clan GH-A, while the GH16 family belonged to clan GH-B. It was observed that GH16 is the largest polyspecific glycoside hydrolase family with ample number of β-agarases and the families GH50 and GH118 were found to be monospecific with only β-agarase activity. And, out of 84 non-catalytic carbohydrate-binding modules (CBMs), only CBM6 and CBM13 were professed in β-agarases. We witnessed a larger heterogeneity in molecular, physiochemical, and catalytic characteristics of the recombinant β-agarases including molecular mass: 32-132 kDa, optimum pH: 4.5-9, optimum temperature 16-60 °C, K M : 0.68-59.8 mg/ml, and V max : 0.781-11,400 U/ mg. Owing to this extensive range of heterogeneity, they have lion's share in the multibillion dollar enzyme market. This review provides a holistic insight to a few aspects of recombinant β-agarases which can be referred by the upcoming explorers to this area.

Expression and Characterization of a Novel β-Porphyranase from Marine Bacterium Wenyingzhuangia fucanilytica: A Biotechnological Tool for Degrading Porphyran

“…The GH16 β-agarases reported after 2010 exhibited different K m values, varying through a wide range from a minimum of 1.82 mg/mL to a maximum value of 59.8 mg/mL. 29 Compared with those enzymes, Por16A_Wf showed a lower K m value, which indicated the desirable binding affinity of the substrate.…”

Algibacillus agarilyticus gen. nov., sp. nov., isolated from the surface of the red algae Gelidium amansii

“…1. 81) are probably responsible for the agar degradation, which breaks down the (l-3)-linked β - d -galactose of agar eclectically and produces neoagarobiose oligosaccharides with d -galactose at the reducing end [24]. Many galactose metabolism pathway key enzymes have been annotated in the genome sequence of strain RQJ05 T (Fig.…”

Characterization of Agarolytic Pathway in a Terrestrial Bacterium Cohnella sp. LGH

“…Thus, the catabolic process of polysaccharides is an important part of global carbon cycle ( Reintjes et al, 2017 ). Due to the biological activities of agaro-oligosaccharides, such as anti-inflammatory ( Kim et al, 2017 ; Ma et al, 2019 ), anti-diabetic ( Park et al, 2020 ), and anti-cariogenic ( Yun et al, 2017 ), agarose is utilized as a raw material in the production of biochemicals ( Veerakumar and Manian, 2018 ; Park et al, 2020 ). Therefore, analysis of the agarose catabolism process by microorganisms not only has significance on f how microorganisms participate in the carbon cycle by utilizing agarose, but also offers a molecular basis for the industrial production of oligomeric sugars and biofuels.…”