title: Purification and characterization of a carboxymethyl cellulase from Artemia salina.
abstract: Brine shrimp (Artemia salina) belong to a group of crustaceans that feed on microalgae and require a cellulase enzyme that can be used in ethanol production from marine algae. Protein with potential cellulase activity was purified and the activity analyzed under different conditions. After initial identification of cellulase activity by CMC cellulase, surface sterilization and PCR using 16s rRNA primers was conducted to confirm that the cellulase activity was not produced from contaminating bacteria. The enzyme was purified by ammonium sulfate fractionation, gel filtration, and ion exchange chromatography. After the final purification, a 70-fold increase in specific enzyme activity was observed. SDS-PAGE results revealed that the cellulase enzyme had a molecular mass of 96 kDa. Temperature, pH, and salinity values were found to be optimal at 55 °C, pH 8.0, and 600 mM NaCl, respectively. Specifically, the enzyme showed a fivefold increase in enzyme activity in seawater compared to 600 mM NaCl in phosphate buffer. Further analysis of the purified enzyme by molecular spectrometry showed no match to known cellulases, indicating this enzyme could be a novel halophilic cellulase that can be used for the production of bioethanol from marine macroalgae.
authors.affiliation: Department of Microbiology, Pukyong National University, 45, Yongso-ro, Nam-Gu, Busan 608-737, Republic of Korea.
authors.forename: Hyun Woo
authors.initials: HW
authors.lastname: Zin
authors.affiliation: Korea Research Institute of Bioscience & Biotechnology (KRIBB), Daejeon 305-806, Republic of Korea.
authors.forename: Kwang-Hyun
authors.initials: KH
authors.lastname: Park
authors.affiliation: Department of Microbiology, Pukyong National University, 45, Yongso-ro, Nam-Gu, Busan 608-737, Republic of Korea. Electronic address: choitj@pknu.ac.kr.
authors.forename: Tae Jin
authors.initials: TJ
authors.lastname: Choi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001156:Artemia / Q000201:enzymology*; D002480:Cellulase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D005591:Chemical Fractionation; D008969:Molecular Sequence Data; D010710:Phosphates / Q000737:chemistry; D054712:Salinity; D012623:Seawater / Q000737:chemistry; D012965:Sodium Chloride
chemical_list: D010710:Phosphates; D012965:Sodium Chloride; D002480:Cellulase; C019826:carboxymethylcellulase
keywords: Artemia; Bioethanol; Cellulase; Halophilic; Macroalgae