Production, Characterization, and Industrial Application of Pectinase Enzyme Isolated from Fungal Strains
Production, Characterization, and Industrial Application of Pectinase Enzyme Isolated from Fungal Strains
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从真菌菌株中分离的果胶酶的生产、表征和工业应用
DOI：
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
V. Raghavan
中科院分区：
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作者：
S. Kc;J. Upadhyaya;D. Joshi;B. Lekhak;Dhiraj Kumar Chaudhary;Bhoj Raj Pant;Tirtha Raj Bajgai;Rajiv Dhital;S. Khanal;Niranjan Koirala;V. Raghavan
果胶酶是一组催化降解果胶物质的酶。它在食品工业中有广泛的应用，用于葡萄酒和果汁的生产和澄清。本研究的目的是从不同土壤样品中分离、筛选和鉴定果胶酶，并评价其在果汁澄清中的应用。用1%柠檬果胶+马铃薯葡萄糖琼脂(PDA)和果胶酶筛选琼脂(PSAM)对果胶分解菌进行初步分离和筛选。采用深层发酵法生产该酶，并用二硝基水杨酸法进行测定。从20个不同的土壤样品中初步筛选出55个真菌菌株，其中只有14个菌株进行了二次筛选。在14个菌株中，只有4个菌株表现出最高的果胶分解活性。4株菌株中，曲霉属(Aspergillusspp.)在培养48h、底物浓度1%、温度30℃时，GM产酶最高。热稳定性测定结果表明，60℃加热10min内，果胶酶活力下降50%，最适温度为30℃，最适pH为5.8U/mL，最适底物浓度为0.5%，最适底物浓度为112.0 U/mL。随着果胶酶浓度的增加，橙汁的出汁率、总可溶性固形物(TSS)和澄清度(%透光率)都有所提高，这表明果胶酶在果汁加工中具有潜在的应用前景。总体而言，曲霉属的菌株。GM被鉴定为一株能够大规模生产果胶酶的菌株。
Pectinases are the group of enzymes that catalyze the degradation of pectic substances. It has wide applications in food industries for the production and clarification of wines and juices. The aim of this study was to isolate, screen and characterize pectinase from fungi isolated from various soil samples and evaluate its application in juice clarification. Fungal strains were isolated and screened primarily using 1% citruspectin incorporated potato dextrose agar (PDA) and secondarily using pectinase screening agar medium (PSAM) for pectinolytic organisms. The enzyme was produced by submerged state fermentation and assayed using the dinitro salicylic acid (DNS) method. From 20 different soil samples, 55 fungal isolates were screened primarily and, among them, only 14 isolates were subjected for secondary screening. Out of 14, only four strains showed the highest pectinolytic activity. Among four strains, Aspergillus spp. Gm showed the highest enzyme production at a 48-h incubation period, 1% substrate concentration, and 30 °C temperature. The thermal stability assessment resulted that the activity of pectinase enzyme declines by 50% within 10 min of heating at 60 °C. The optimum temperature, pH, and substrate concentration for the activity of enzyme was 30 °C (75.4 U/mL), 5.8 (72.3 U/mL), and 0.5% (112.0 U/mL), respectively. Furthermore, the yield of the orange juice, the total soluble solid (TSS), and clarity (% transmittance) was increased as the concentration of the pectinase increased, indicating its potential use in juice processing. Overall, the strain Aspergillus spp. Gm was identified as a potent strain for pectinase production in commercial scale.