title: High production of cold-tolerant chitinases on shrimp wastes in bench-top bioreactor by the Antarctic fungus Lecanicillium muscarium CCFEE 5003: bioprocess optimization and characterization of two main enzymes.
abstract: The Antarctic fungus Lecanicillium muscarium CCFEE-5003 was preliminary cultivated in shaken flasks to check its chitinase production on rough shrimp and crab wastes. Production on shrimp shells was much higher than that on crab shells (104.6±9.3 and 48.6±3.1U/L, respectively). For possible industrial applications, bioprocess optimization was studied on shrimp shells in bioreactor using RSM to state best conditions of pH and substrate concentration. Optimization improved the production by 137% (243.6±17.3). Two chitinolytic enzymes (CHI1 and CHI2) were purified and characterized. CHI1 (MW ca. 61kDa) showed optima at pH 5.5 and 45°C while CHI2 (MW ca. 25kDa) optima were at pH 4.5 and 40°C. Both enzymes maintained high activity levels at 5°C and were inhibited by Fe(++), Hg(++) and Cu(++). CHI2 was strongly allosamidin-sensitive. Both proteins were N-acetyl-hexosaminidases (E.C. 3.2.1.52) but showed different roles in chitin hydrolysis: CHI1 could be defined as "chitobiase" while CHI2 revealed a main "eso-chitinase" activity.
authors.affiliation: Dipartimento di Scienze Ecologiche e Biologiche, Agroalimentari e Forestali, Largo Università snc, University of Tuscia, I-01100 Viterbo, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Barghini
authors.forename: Deborah
authors.initials: D
authors.lastname: Moscatelli
authors.forename: Anna Maria Vittoria
authors.initials: AM
authors.lastname: Garzillo
authors.forename: Silvia
authors.initials: S
authors.lastname: Crognale
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Fenice
mesh_terms: D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D000864:Antarctic Regions; D057005:Bioengineering; D019149:Bioreactors / Q000382:microbiology*; D003386:Brachyura; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000096:biosynthesis* / Q000737:chemistry; D003080:Cold Temperature; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005285:Fermentation; D006863:Hydrogen-Ion Concentration; D006999:Hypocreales / Q000201:enzymology* / Q000302:isolation & purification; D008970:Molecular Weight; D033561:Penaeidae; D012623:Seawater / Q000382:microbiology; D014312:Trisaccharides / Q000494:pharmacology; D014866:Waste Products / Q000032:analysis
chemical_list: D004791:Enzyme Inhibitors; D014312:Trisaccharides; D014866:Waste Products; C054851:allosamidin; D002686:Chitin; D002688:Chitinases; D000117:Acetylglucosamine
keywords: Biochemical characterization; Cold-tolerant chitinolytic enzymes; Lecanicillium muscarium; RSM optimization; Shrimp and crab wastes