Isolasi, Skrining, dan Identifikasi Molekuler Bakteri Termotoleran Proteolitik dari Sumber Air Panas Nglimut Gonoharjo Kendal

Abstract

The potential for the use of enzymes in industry is starting to be widely used. Protease is one of the most widely used enzymes in industry. Protease is an enzyme capable of hydrolyzing peptide bonds in proteins. Proteolytic thermotolerant bacteria are one of the important sources of thermostable enzymes that can be isolated from hot springs. The purpose of this study was to isolate and screen proteolytic thermotolerant bacteria and to identify molecularly the best isolates of proteolytic thermotolerant bacteria from the hot springs of Nglimut Gonoharjo. Isolation was carried out by graded dilution on Sodium Agar (NA) and Thermus Agar (TA) media using the spread plate method, and the proteolytic activity test using the paper disc method. Characterization of proteolytic bacterial isolates was carried out by macroscopic and microscopic morphology with gram staining. Molecular identification with 16S rRNA gene through amplification, electrophoresis, sequencing, BLAST analysis and phylogenetic reconstruction. The isolation results obtained 32 proteolytic thermotolerant isolates with irregular and round colony shape characteristics, Gram positive and Gram negative, and stem cells. The results of the enzyme test showed that isolate T5 was the best isolate with a proteolytic index value of 1.90 and was identified as Bacillus subtilis. Gram Positive and Gram Negative, and have stem cells. The results of the enzyme test showed that isolate T5 was the best isolate with a proteolytic index value of 1.90 and was identified as Bacillus subtilis.