Abstract
Napa cabbage waste contains an organic component, cellulose, which can be utilised as an ingredient for cellulose-degrading enzyme production with the help of indigenous yeast. The aim of the research was to identify and characterise potential indigenous yeast isolated from napa cabbage waste, which has cellulose-degrading activity. Indigenous yeast were isolated and characterised using the RapID Yeast Plus System, then turbidity was used to determine the yeast total population. Indigenous yeast was grown at napa cabbage waste at 27, 37, and 40°C for three days, and cellulose-degrading activity was determined by the Dinitrosalicylic Acid (DNS) method. The potential yeast isolate with the highest cellulose-degrading activity was identified by a sequence analysis of the rRNA gene internal transcribed spacer (ITS) region with using primers ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′- TCCTCCGCTTATTGATATGC-3′). The results were compared to the GenBank database using the Basic Local Alignment Search Tools/BLAST algorithm. Three species of indigenous yeast were isolated from napa cabbage waste (S2, S6, and S8). S8, incubated at 37ºC for three days, demonstrated the highest cellulose-degrading enzyme activity (1.188 U/mL), with the average activity of 0.684U/mL. Species identification results indicated that the S8 isolate had a 100% similarity to Pichia fermentans UniFGPF2 (KT029805.1).
Highlights
Napa cabbage (Brassica pekinesis L.) is one of the most cultivated agricultural products in Indonesia
The isolation of indigenous cellulolytic yeast from napa cabbage waste was carried out using the direct plating method [5,6]
One gram of crushed napa cabbage waste was added into 0.85% NaCl, inoculated into a modified PDA (PDA with a 3% yeast extract and 10 ppm antibiotics) and incubated at 30°C for three days
Summary
Napa cabbage (Brassica pekinesis L.) is one of the most cultivated agricultural products in Indonesia. Napa cabbage waste is used as animal feed, while its value could be increased, e.g., through production of cellulose-degrading enzymes. The fact that temperature is an controlled parameter makes it possible to support yeast growth during the fermentation of napa cabbage waste for cellulose-degrading enzyme production. The selection of the best treatment was performed cellulose-degrading enzymes in supernatant, where based on quantitative analysis by determining the crude enzymes reacted with DNS (Dinitrosalicylic highest value of enzyme activity using Factorial Acid) reagent. According to the results of carried out to obtain absorbance values which were used isolation and identification of indigenous cellulolytic to determine cellulose-degrading enzyme activity. The yeast from napa cabbage waste, descriptive analysis on control used was 3 mL of DNS reagent that was diluted the total population of yeast during the production of to 25 mL by distilled water.
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