Identification of extracellular enzyme producing alkalophilic bacilli from Izmir province by 16S-ITS rDNA RFLP.
OBJECTIVE
To screen industrially important extracellular enzymes from the newly isolated alkalophilic bacilli and to characterize them by phenotypic and 16S-internal transcribed spacer (ITS) rDNA restriction pattern analysis.


RESULTS
Three different environmental samples, soil, leather and horse faeces, were collected within the province of Izmir. Isolates grown on Horikoshi-I medium for 24 h at 37 degrees C were screened for extracellular enzyme activity by using eight different substrates: birchwood xylan, carboxymethylcellulose, casein, citrus pectin, polygalacturonic acid, soluble starch, and Tween 20 and 80. In total, 115 extracellular enzyme-producing bacilli were obtained. Casein was hydrolysed by 78%, soluble starch by 67%, citrus pectin by 63%, polygalacturonic acid by 62%, Tween 20 by 34%, birchwood xylan by 16%, Tween 80 by 12%, and carboxymethylcellulose by 3% of the isolates. The isolates were differentiated into 19 distinct homology groups by the 16S-ITS rDNA restriction pattern analysis.


CONCLUSIONS
Eight different extracellular enzyme activities were determined in 115 endospore forming bacilli. The largest 16S-ITS rDNA homology group (HT1) included 36% of the isolates, 98% of which degraded casein, polygalacturonic acid, pectin and starch.


CONCLUSIONS
This study is the first report on the characterization of the industrial enzyme-producing alkalophilic bacilli by 16S-ITS rDNA restriction fragment length polymorphism (RFLP). Restriction profiles of 64% of the isolates were found to be different from those of five reference strains used.
Akbalik
G
G
Biotechnology and Bioengineering Programme, Science Faculty, Izmir Institute of Technology, Gulbahce, Izmir, Turkey.
Gunes
H
H
Yavuz
E
E
Yasa
I
I
Harsa
S
S
Elmaci
Z S
ZS
Yenidunya
A F
AF
D000818:Animals; D001407:Bacillus / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D002364:Caseins / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005243:Feces / Q000382:microbiology; D006736:Horses; D006863:Hydrogen-Ion Concentration; D010368:Pectins / Q000378:metabolism; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D012689:Sequence Homology, Nucleic Acid; D012988:Soil Microbiology; D013213:Starch / Q000378:metabolism; D013696:Temperature; D014421:Turkey
D002364:Caseins; D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; D010368:Pectins; D013213:Starch; D015252:Deoxyribonucleases, Type II Site-Specific; C024403:GGCC-specific type II deoxyribonucleases; C027046:TCGA-specific type II deoxyribonucleases; D010447:Peptide Hydrolases; C003181:polygalacturonic acid