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Sebastian Margino
Gadjah Mada University
Agriculture, Biological Sciences & Forestry

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Protoplast Fusion of Koji-Mold Aspergillus spp. to Improve Kecap Production Sebastian Margino; F. S. Rejeki; E. S. Rahayu; D. Wibowo
Indonesian Food and Nutrition Progress Vol 5, No 1 (1998)
Publisher : Indonesian Association of Food Technologists

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jifnp.60

Abstract

Protoplast fusion of Aspergillus oryzae and A. niger for improving the enzyme productivity of desirable kojimolds for kecap (Indonesian soy sauce) was studied. Protoplasting of both mycellium aspergilli was obtained by treating them using Novozyme 234, at final concen tration 5 mg/ml and the time of exposure for three and two hours. Regenerated protoplasts of A. oryzae oncomplete medium (RCMMPA) was 1.46-3.74 % and on minimal medium (RMMPA) was 0.93-4.93%, whereas A. niger was 0.44-7.64 % and 0.17-6.25 %, respectively. Protoplast fusion was conducted in 30 % PEG 6000 (Polyethylene Glycol) containing 0.01 M CaC12 and 0.05 % glycine, and at pH 7.5. For selecting the suspected fusant Dinikonasol 12.5%, at the final concentration 200 ppm and Cycloheximide at the final concentration 100 ppm Was used as a marker. Results showed that hundred eighteen of suspected fusants were obtained and after screening found that seven as selected fusants. Based on the enzyme activities, finally, fusant number 77 was selected as a fusant with high proteolytic and glutaminase activities.
Mutation Technique for Increasing the Production of Antibacteria Lactobacillus plantarum TGR-2 Sebastian Margino; Sri Winarti; Retno Indrati; Endang S. Rahayu
Indonesian Food and Nutrition Progress Vol 5, No 2 (1998)
Publisher : Indonesian Association of Food Technologists

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jifnp.71

Abstract

Production of antibacteria of Lactobacillus plantarum TGR-2 was carried out by mutation technique using UV radiation and chemical mutagenic agent (acrydine orange (AO), ethyl methane sulfonate (EMS), and N-methyl-N-nitro sulfonate nitrosoguanidine (NTG). As a marker was applied chloramphenicol for 10 ppm. Total 214 mutants were obtained from all of the treatments and then screened based on their activities against Staphylococcus aureus FNCC 0047 as bacterial indicator, using agar diffusion and turbidimetric assay techniques. Results showed that 24 mutants could produce higher amount of antibacteria and two of them, NTG and acrydine orange treatments, had higher stability than others. Characterization of two mutants, determination of biochemistry traits and optimization of antibacterial (bacteriocin) production were done. The product showed that UV-15, EMS-10, AO-6 and NTG-21 mutants could produce antibacterial substance 3,90, 4.40, 3.40 and 4.17 AU/ml for 16, 12, 12, and 16 hours incubation, respectively, compared to Lactobacillus plantarum TGR-2 which produced 1.07 UA/ ml for 24 hours incubation.
Co-Authors D. Wibowo E. S. Rahayu Endang S. Rahayu F. S. Rejeki Retno Indrati Sri Winarti