Thermoactive extracellular proteases of Geobacillus caldoproteolyticus, sp. nov., from sewage sludge
Tóm tắt
A proteolytic thermophilic bacterial strain, designated as strain SF03, was isolated from sewage sludge in Singapore. Strain SF03 is a strictly aerobic, Gram stain-positive, catalase-positive, oxidase-positive, and endospore-forming rod. It grows at temperatures ranging from 35 to 65°C, pH ranging from 6.0 to 9.0, and salinities ranging from 0 to 2.5%. Phylogenetic analyses revealed that strain SF03 was most similar to Saccharococcus thermophilus, Geobacillus caldoxylosilyticus, and G. thermoglucosidasius, with 16S rRNA gene sequence identities of 97.6, 97.5 and 97.2%, respectively. Based on taxonomic and 16S rRNA analyses, strain SF03 was named G. caldoproteolyticus sp. nov. Production of extracellular protease from strain SF03 was observed on a basal peptone medium supplemented with different carbon and nitrogen sources. Protease production was repressed by glucose, lactose, and casamino acids but was enhanced by sucrose and NH4Cl. The cell growth and protease production were significantly improved when strain SF03 was cultivated on a 10% skim-milk culture medium, suggesting that the presence of protein induced the synthesis of protease. The protease produced by strain SF03 remained active over a pH range of 6.0–11.0 and a temperature range of 40–90°C, with an optimal pH of 8.0–9.0 and an optimal temperature of 70–80°C, respectively. The protease was stable over the temperature range of 40–70°C and retained 57 and 38% of its activity at 80 and 90°C, respectively, after 1 h.
Tài liệu tham khảo
Ahmad S, Scopes RK, Rees GN, Patel BKC (2000) Saccharococcus caldoxylosilyticus sp. nov., an obligately thermophilic, xylose-utilizing, endospore-forming bacterium. Int J Syst Evol Microbiol 50:517–523
Antranikian G, Klingeberg M (1991a) Thermostable protease from Thermococcus. PCT Patent Appl WO 9119792
Antranikian G, Klingeberg M (1991b) Thermostable protease from Staphylothermus. PCT Patent Appl WO 9119791
Anwar A, Saleemuddin M (1998) Alkaline proteases: a review. Bioresource Technol 64:175–183
Ash C, Farrow JAE, Wallbanks S, Collins MD (1991) Phylogenetic heterogeneity of the genus Bacillus revealed by comparative analysis of small-subunit-ribosomal RNA sequences. Lett Appl Microbiol 13:202–206
Boethling RS (1975) Regulation of extracellular protease secretion in Pseudomonas maltophilia. J Bacteriol 123:954–961
Cowan DA, Smolenski KA, Daniel RM, Morgan HW (1987) An extremely thermostable extracellular proteinase from a strain of the archaebacterium Desulfurococcus growing at 88°C. Biochem J 247:121–133
Denkin SM, Nelson DR (1999) Induction of protease activity in Vibrio anguillarum by gastrointestinal mucus. Appl Environ Microbiol 65:3555–3560
Dhandapani R, Vijayaragavan R (1994) Production of a thermophilic extracellular alkaline protease by Bacillus stearothermophilus AP-4. World J Microbiol Biotechnol 10:33–35
Eggen R, Geerling A, Watts J, de Vos WM (1990) Characterization of pyrolysin, a hyperthermoactive serine protease from the archaeobacterium Pyrococcus furiosus. FEMS Microbiol Lett 71:17–20
Felsenstein J (1989) PHYLIP (PHYlogenetic Inference Package). Cladistics 5:164–166
Ferrero MA, Castro GR, Abate CM, Baigorí MD, Siñeriz F (1996) Thermostable alkaline proteases of Bacillus licheniformis MIR 29: isolation, production and characterization. Appl Microbiol Biotechnol 45:327–332
Fortina MG, Mora D, Schumann P, Parini C, Manachini PL, Stackebrandt E (2001) Reclassification of Saccharococcus caldoxylosilyticus as Geobacillus caldoxylosilyticus (Ahmad et al. 2000) comb. nov. Int J Syst Evol Microbiol 51:2063–2071
Godfrey T and West S (1996) Introduction to industrial enzymology. In: Godfrey T, West S (eds) Industrial enzymology, 2nd edn. Macmillan, London, pp 1–8
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98
Heinen UJ, Heinen W (1972) Characteristics and properties of a caldoactive bacterium producing extracellular enzymes and two related strains. Arch Mikrobiol 82:1–23
Kalisz HM (1988) Microbial proteinases. Adv Biochem Eng Biotechnol 36:1–65
Kumar CG, Takagi H (1999) Microbial alkaline proteases: from a bioindustrial viewpoint. Biotechnol Adv 17:561–594
Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. Wiley, Chichester, pp 115–175
Lee JK, Kim YO, Kim HK, Park YS, Oh TK (1996) Purification and characterization of a thermostable alkaline protease from Thermoactinomyces sp. E79 and the DNA sequence of the encoding gene. Biosci Biotechnol Biochem 60:840–846
Maidak BL, Cole JR, Lilburn TG, Parker CT Jr, Saxman PR, Stredwick JM, Garrity GM, Li B, Olsen GJ, Pramanik S, Schmidt TM, Tiedje JM (2000) The RDP (Ribosomal Database Project) continues. Nucleic Acids Res 28:173–174
Manachini PL, Mora D, Nicastro G, Parini C, Stackebrandt E, Pukall R, Fortina MG (2000). Bacillus thermodenitrificans sp. nov., nom. rev. Int J Syst Evol Microbiol 50:1331–1337
Matsuzawa H, Hamaoki M, Ohta T (1983) Production of thermophilic extracellular proteases (aqualysins I and II) by Thermus aquaticus YT-1. Agric Biol Chem 47:25–28
Morikawa M, Izawa Y, Rashid N, Hoaki T, Imanaka T (1994) Purification and characterization of a thermostable thiol protease from a newly isolated hyperthermophilic Pyrococcus sp. Appl Environ Microbiol 60:4559–4566
Nazina TN, Tourova TP, Poltaraus AB, Novikova EV, Grigoryan AA, Ivanova AE, Lysenko AM, Petrunyaka VV, Osipov GA, Belyaev SS, Ivanov MV (2001) Taxonomic study of aerobic thermophilic bacilli: descriptions of Geobacillus subterraneus gen. nov., sp. nov. and Geobacillus uzenensis sp. nov. from petroleum reservoirs and transfer of Bacillus stearothermophilus, Bacillus thermocatenulatus, Bacillus thermoleovorans, Bacillus kaustophilus, Bacillus thermoglucosidasius and Bacillus thermodenitrificans to Geobacillus as the new combinations G. stearothermophilus, G. thermocatenulatus, G. thermoleovorans, G. kaustophilus, G. thermoglucosidasius and G. thermodenitrificans. Int J Syst Evol Microbiol 51:433–446
Nystrand R (1984) Saccharococcus thermophilus gen. nov., sp. nov., isolated from beet sugar extraction. Syst Appl Microbiol 5:204–219
Peek K, Daniel RM, Monk C, Parker L, Coolbear T (1992) Purification and characterization of a thermostable proteinase isolated from Thermus sp. strain Rt41A. Eur J Biochem 207:1035–1044
Rahman RNZA, Razak CN, Ampon K, Basri M, Yunus WMZW, Salleh AB (1994) Purification and characterization of a heat-stable alkaline protease from Bacillus stearothermophilus F1. Appl Microbiol Biotechnol 40:822–827
Rainey FA, Stackebrandt E (1993) Phylogenetic evidence for the relationship of Saccharococcus thermophilus to Bacillus stearothermophilus. Syst Appl Microbiol 16:224–226
Rainey FA, Fritze D, Stackebrandt E (1994). The phylogenetic diversity of thermophilic members of the genus Bacillus as revealed by 16S rDNA analysis. FEMS Microbiol Lett 115:205–211
Secades P, Guijarro JA (1999) Purification and characterization of an extracellular protease from the fish pathogen Yersinia ruckeri and effect of culture conditions on production. Appl Environ Microbiol 65: 3969–3975
Smiber RM, Krieg NR (1994) Phenotypic characterization. In: Gerhardt P, Murray RGE, Wood WA, Krieg NR (eds) Methods for general and molecular bacteriology. American Society for Microbiology, Washington, pp 607–654
Sneath PHA (1986) Endospore-forming Gram-positive rods and cocci. In: Sneath PHA, Mair NS, Sharp ME, Holt JG (eds) Bergey’s manual of systematic bacteriology, vol 2. Williams and Wilkins, Baltimore, pp 1104–1139
Stackebrandt E, Goebel BM (1994) Taxonomic note: a place for DNA–DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol 44:846–849
Sunna A, Tokajian S, Burghardt J, Rainey F, Antranikian G, Hashwa F (1997) Identification of Bacillus kaustophilus, Bacillus thermocatenulatus and Bacillus strain HSP as members of Bacillus thermoleovorans. Syst Appl Microbiol 20:232–237
Suzuki Y, Kishigami T, Inoue K, Mizoguchi Y, Eto N, Takagi M, Abe S (1983) Bacillus thermoglucosidasius sp. nov., a new species of obligately thermophilic bacilli. Syst Appl Microbiol 4:487–495
Tsuchiya K, Nakamura Y, Sakashita H, Kimura T (1992) Purification and characterization of thermostable alkaline protease from alkalophilic Thermoactinomyces sp HS682. Biosci Biotechnol Biochem 56:246–250
Ursing JB, Rosselló-Mora RA, García-Valdés E, Lalucat J (1995) Taxonomic note: a pragmatic approach to the nomenclature of phenotypically similar genomic groups. Int J Syst Bacteriol 45:604
Voordouw G, Roche RS (1975) The role of bound calcium ions in thermostable, proteolytic enzymes. II. Studies on thermolysin, the thermostable protease from Bacillus thermoproteolyticus. Biochemistry 14:4667–4673
Yoshikawa M, Matsuda F, Naka M, Murofushi E, Tsunematsu. J (1974) Pleiotropic alterations of activities of several toxins and enzymes in mutants of Staphylococcus aureus. J Bacteriol 119:117–122