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Optimization of Trypsin-like Protease Production by Lactobacillus plantarum FNCC 0270 using Response Surface Methodology Trismilah,; Nurhasanah, Astutiati; Sumaryono, Wahono; Malik, Amarila; Sadikin, Mohamad
Makara Journal of Science Vol. 19, No. 2
Publisher : UI Scholars Hub

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The purpose of this study was to get optimum medium composition and agitation to trypsin-like protease production by Lactobacillus plantarum FNCC 0270. The medium composition and agitation for enzyme production was optimized using Central Composite Design and Response Surface Method with Design Expert software version 7.1.5 Fermentation was carried out in erlenmeyer flasks at initial pH 8, 37 °C, using an incubator shaker at 87.5 rpm. The best results showed an enzyme activity of 1.0 mU/mL, a protein level of 0.557 mg/mL, and desirability value of 0.740. Numerical optimization was performed to approach the ideal state of the fermentation or the desirability value of 1. The medium composition containing of 3.64% baker's yeast, 1.21% glucose, and 0.13% skim milk was used for the fermentation. The enzyme activity of 1.51 mU/mL and protein level of 0.205 mg/mL can be achieved. After numerical optimization, the fermentation process was verified in erlenmeyer flasks with incubator shaking at 77 rpm, initial pH 8, 37 °C, and 15 h fermentation. The verification results showed that the enzyme activity of 1.273 ± 0.227 mU/mL and protein level of 0.248 ± 0.012 mg/mL.

PRODUKSI BETA-1,3 GLUKAN DARI Agrobacterium DAN AKTIVITAS PENYEMBUHAN LUKA TERBUKA PADA TIKUS PUTIH Kusmiati, Kusmiati; Rachmawati, Fitria; Siregar, Syafrida; Nuswantara, Sukma; Malik, Amarila
Makara Journal of Science Vol. 10, No. 1
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Production of beta-1,3 glucan from Agrobacterium and its wound healing activity on white rat. The objective of this study was to determine the activity of beta-1,3 glucan product extracted from local Agrobacterium sp Bro 1.2.1, both wild-type and mutant-type, on opened-wound healing process. Beta-1,3 glucan product was extracted by precipitation, and the purification was carried out by column chromatography as KCl gradient fractions. In this study, white Sprague Dawley rats were employed, and have been treated for opened-wound condition. Seven groups were performed in this experiment, i.e. the negative control, the positive control employing povidone iodine, the two groups of two commercial beta-1,3 glucan with 0,02 mg/4 cm2 each, and the last three groups of beta-1,3 glucan as the test group with 0,02 mg/4 cm2, 0,10 mg/4 cm2 and 0,50 mg/4 cm2, respectively. The result showed significant differences of wound-healing activity performing statistical analysis of the least significance between the negative control, the positive control, as well as the highest dose of the test group of beta-1,3 glucan, at the dose of 0,5 mg/4 cm2 (p<0.05).

PRODUKSI BETA-1,3 GLUKAN DARI Agrobacterium DAN AKTIVITAS PENYEMBUHAN LUKA TERBUKA PADA TIKUS PUTIH Kusmiati, Kusmiati; Rachmawati, Fitria; Siregar, Syafrida; Nuswantara, Sukma; Malik, Amarila
Makara Journal of Science Vol. 10, No. 1
Publisher : UI Scholars Hub

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Production of beta-1,3 glucan from Agrobacterium and its wound healing activity on white rat. The objective of this study was to determine the activity of beta-1,3 glucan product extracted from local Agrobacterium sp Bro 1.2.1, both wild-type and mutant-type, on opened-wound healing process. Beta-1,3 glucan product was extracted by precipitation, and the purification was carried out by column chromatography as KCl gradient fractions. In this study, white Sprague Dawley rats were employed, and have been treated for opened-wound condition. Seven groups were performed in this experiment, i.e. the negative control, the positive control employing povidone iodine, the two groups of two commercial beta-1,3 glucan with 0,02 mg/4 cm2 each, and the last three groups of beta-1,3 glucan as the test group with 0,02 mg/4 cm2, 0,10 mg/4 cm2 and 0,50 mg/4 cm2, respectively. The result showed significant differences of wound-healing activity performing statistical analysis of the least significance between the negative control, the positive control, as well as the highest dose of the test group of beta-1,3 glucan, at the dose of 0,5 mg/4 cm2 (p<0.05).

PRODUKSI BETA-1,3 GLUKAN DARI Agrobacterium DAN AKTIVITAS PENYEMBUHAN LUKA TERBUKA PADA TIKUS PUTIH Kusmiati, Kusmiati; Rachmawati, Fitria; Siregar, Syafrida; Nuswantara, Sukma; Malik, Amarila
Makara Journal of Science Vol. 10, No. 1
Publisher : UI Scholars Hub

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Production of beta-1,3 glucan from Agrobacterium and its wound healing activity on white rat. The objective of this study was to determine the activity of beta-1,3 glucan product extracted from local Agrobacterium sp Bro 1.2.1, both wild-type and mutant-type, on opened-wound healing process. Beta-1,3 glucan product was extracted by precipitation, and the purification was carried out by column chromatography as KCl gradient fractions. In this study, white Sprague Dawley rats were employed, and have been treated for opened-wound condition. Seven groups were performed in this experiment, i.e. the negative control, the positive control employing povidone iodine, the two groups of two commercial beta-1,3 glucan with 0,02 mg/4 cm2 each, and the last three groups of beta-1,3 glucan as the test group with 0,02 mg/4 cm2, 0,10 mg/4 cm2 and 0,50 mg/4 cm2, respectively. The result showed significant differences of wound-healing activity performing statistical analysis of the least significance between the negative control, the positive control, as well as the highest dose of the test group of beta-1,3 glucan, at the dose of 0,5 mg/4 cm2 (p<0.05).

ISOLASI DAN SKRINING MOLEKULER BAKTERI ASAM LAKTAT PEMBAWA GEN GLUKANSUKRASE DARI MAKANAN DAN MINUMAN MENGANDUNG GULA Malik, Amarila; Hermawati, Ajitya Kurnia; Hestiningtyas, Mahardhika; Radji, Maksum
Makara Journal of Science Vol. 14, No. 1
Publisher : UI Scholars Hub

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Isolation and Molecular Screening of Glucansucrase Gene Harboring-Lactic Acid Bacteria. Exopolysaccharides (EPS) have been possessed to be used in pharmaceutical, cosmetic and food industries. Lactic acid bacteria (LAB) produce a wide variety of exopolysaccharides and carries sucrase genes glucansucrase/glucosyltransferase (gtf) and fructansucrase/fructosyltransferases (ftf) for EPS production. In this study, the isolation and screening of EPS producing-LAB (EPS-LAB) were carried out on modified de Mann-Rogosa-Sharpe (MRS) agar medium supplemented with 10% of sucrose on LAB isolated from various unique sugar containing-foods and -beverages originated from local sources. Besides obtaining EPS-LAB, this study aimed to screen for gtf gene as well as to molecular identify strains by using PCR technique. Degenerate primer pairs DegFor and DegRev which targeted the conserved region of gtf genes catalytic domain were used, whereas LABfw and LABrv were used to molecular identify strains using 16S rRNA gene. An approximately 660 base pairs (bp) amplicons which targeted gtf gene were obtained from 13 out of 16 srains chosen. Result from PCR of 16S rRNA gene identification on gtf positive strains revealed that all strains were molecular identified as LAB after DNA sequencing analysis by using blastn. A rare EPS-producing LAB were obtain i.e. Weissella. Strains obtained in this study are potential sources for exploring novel sucrase gene/s and obtain unique EPS polymer product/s.

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