title: Efficient enzymatic degradation process for hydrolysis activity of the Carrageenan from red algae in marine biomass.
abstract: Carrageenan is a generic name for a family of polysaccharides obtained from certain species of red algae. New methods to produce useful cost-efficiently materials from red algae are needed to convert enzymatic processes into fermentable sugars. In this study, we constructed chimeric genes cCgkA and cCglA containing the catalytic domain of κ-carrageenase CgkA and λ-carrageenase CglA from Pseudoalteromonas carrageenovora fused with a dockerin domain. Recombinant strains expressing the chimeric carrageenase resulted in a halo formation on the carrageenan plate by alcian blue staining. The recombinant cCgkA and cCglA were assembled with scaffoldin miniCbpA via cohesin and dockerin interaction. Carbohydrate binding module (CBM) in scaffoldin was used as a tag for cellulose affinity purification using cellulose as a support. The hydrolysis process was monitored by the amount of reducing sugar released from carrageenan. Interestingly, these results indicated that miniCbpA, cCgkA and cCglA assembled into a complex and that the dockerin-fused enzymes on the scaffoldin had synergistic activity in the degradation of carrageenan. The observed enhancement of activity by carrageenolytic complex was 3.1-fold-higher compared with the corresponding enzymes alone. Thus, the assemblies of advancement of active enzyme complexes will facilitate the commercial production of useful products from red algae biomass which represents inexpensive and sustainable feed-stocks.
authors.affiliation: Department of Biotechnology, Korea University, Seoul 136-701, Republic of Korea.
authors.forename: Dae Hee
authors.initials: DH
authors.lastname: Kang
authors.affiliation: Department of Biotechnology, Korea University, Seoul 136-701, Republic of Korea.
authors.forename: Jeong Eun
authors.initials: JE
authors.lastname: Hyeon
authors.affiliation: Department of Biotechnology, Korea University, Seoul 136-701, Republic of Korea.
authors.forename: Seung Kyou
authors.initials: SK
authors.lastname: You
authors.affiliation: Department of Chemical and Biological Engineering, Korea University, Seoul 136-701, Republic of Korea.
authors.forename: Seung Wook
authors.initials: SW
authors.lastname: Kim
authors.affiliation: Department of Biotechnology, Korea University, Seoul 136-701, Republic of Korea. Electronic address: samhan@korea.ac.kr.
authors.forename: Sung Ok
authors.initials: SO
authors.lastname: Han
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018533:Biomass; D002351:Carrageenan / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D006868:Hydrolysis; D031161:Pseudoalteromonas / Q000201:enzymology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D000461:Rhodophyta; D000097722:Cohesins
chemical_list: D001426:Bacterial Proteins; D002351:Carrageenan; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D006026:Glycoside Hydrolases; D011993:Recombinant Fusion Proteins; D000097722:Cohesins
keywords: Carbohydrate binding module; Carrageenase; Cohesin; Dockerin; Scaffoldin