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      Shewanella algae and Microbulbifer elongatus from marine macro-algae – isolation and characterization of agar-hydrolysing bacteria

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          Abstract

          Macro-algae are a good source of agar oligosaccharides, which can be obtained through bacterial enzymatic hydrolysis. The agarase enzyme secreted by the micro-organisms cleaves the cell wall of the algae and releases agar oligosaccharides as degradation products with various applications. Agarolytic bacteria were isolated from the marine algae Kappaphycus sp., and Sargassum sp., and studied for their agar-degrading properties. Among the 70 isolates, 2 isolates (A13 and Sg8) showed agarase activity in in vitro assays. The maximum agarolytic index was recorded in the isolate Sg8 (3.75 mm and 4.29 µg ml −1 agarase activity), followed by the isolate A13 (2.53 mm and 2.6 µg ml −1 agarase activity). Optimum agarase production of isolate Sg8 was observed at pH7 and at a temperature of 25 °C in 24–48 h, whereas for isolate A13 the optimum production was at pH7 and at a temperature of 37 °C in 48 h. The identities of the agarolytic isolates (Sg8 and A13) were confirmed based on microscopy, morphological, biochemical and molecular analysis as Shewanella algae [National Center for Biotechnology Information (NCBI) GenBank accession number MK121204.1] and Microbulbifer elongatus [NCBI GenBank accession number MK825484.1], respectively.

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          Measurement of carboxymethylcellulase activity

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            Agarase: Review of Major Sources, Categories, Purification Method, Enzyme Characteristics and Applications

            Agarases are the enzymes which catalyze the hydrolysis of agar. They are classified into α-agarase (E.C. 3.2.1.158) and β-agarase (E.C. 3.2.1.81) according to the cleavage pattern. Several agarases have been isolated from different genera of bacteria found in seawater and marine sediments, as well as engineered microorganisms. Agarases have wide applications in food industry, cosmetics, and medical fields because they produce oligosaccharides with remarkable activities. They are also used as a tool enzyme for biological, physiological, and cytological studies. The paper reviews the category, source, purification method, major characteristics, and application fields of these native and gene cloned agarases in the past, present, and future.
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              Cloning, expression, and characterization of a glycoside hydrolase family 86 beta-agarase from a deep-sea Microbulbifer-like isolate.

              The gene for a novel beta-agarase from a deep-sea Microbulbifer-like isolate was cloned and sequenced. It encoded a mature protein of 126,921 Da (1146 amino acids), which was a modular protein including two tandem carbohydrate-binding module (CBM)-like sequences and a catalytic module. The catalytic module resembled a glycoside hydrolase family 86 beta-agarase, AgrA, from Pseudoalteromonas atlantica T6c with 31% amino acid identity. Its recombinant agarase was hyper-produced extracellularly using Bacillus subtilis as the host and purified to homogeneity. The activity and stability were strongly enhanced by CaCl2. The maximal enzyme activity was observed at 45 degrees C and pH 7.5 in the presence of 10 mM CaCl2. The enzyme was an endo-type beta-agarase and degraded agarose and agarose oligosaccharides more polymerized than hexamers to yield neoagarohexaose as the main product. This is the first glycoside hydrolase family 86 enzyme to be homogeneously purified and characterized.
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                Author and article information

                Journal
                Access Microbiol
                Access Microbiol
                acmi
                acmi
                Access Microbiology
                Microbiology Society
                2516-8290
                2020
                10 September 2020
                10 September 2020
                : 2
                : 11
                : acmi000170
                Affiliations
                [ 1] T. Stanes and Company Limited , Coimbatore, India
                Author notes

                The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA sequences of Shewanella algae and Microbulbifer elongatus are MK121204.1 and MK825484.1, respectively.

                [†]

                These authors contributed equally to this work

                [‡]

                These authors also contributed equally to this work

                Author information
                https://orcid.org/0000-0001-8456-0512
                Article
                000170
                10.1099/acmi.0.000170
                7717482
                33294773
                0c1c1d98-ede3-4e14-acad-999a215296a0
                © 2020 The Authors

                This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License.

                History
                : 08 November 2019
                : 24 August 2020
                Funding
                Funded by: BIRAC
                Award ID: BT/SBIRI/1394/31/16
                Award Recipient : Latha K
                Categories
                Short Communication
                Biotechnology
                Other Bacteria
                Custom metadata
                0

                agar-hydrolysing bacteria,kappaphycus sp.,marine macro-algae,microbulbifer elongatus,sargassum sp.,shewanella algae

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