果胶酶粗酶液与漆酶粗酶液复配酶解膨胀梗丝的工艺优化
Optimization of enzymatic hydrolysis process for expanded tobacco cut stem using combined crude enzymes of pectinase and laccase
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摘要: 对从烟田土壤和腐朽烟梗中筛选获得产果胶酶菌株TS-1和产漆酶菌株TS-2进行形态学观察和分子生物学鉴定,通过单因素试验优化二者的产酶条件,并将果胶酶粗酶液与漆酶粗酶液复配酶解膨胀梗丝,研究膨胀梗丝感官品质及致香成分的变化。结果表明:菌株TS-1和TS-2分别鉴定为黑曲霉(Aspergillus niger)和黄孢原毛平革菌(Phanerodontia chrysosporium),二者最优产果胶酶和漆酶条件分别为烟梗粉质量浓度50 g/L、发酵温度30 ℃、发酵pH值6.0及烟梗粉质量浓度50 g/L、(NH4)2SO4质量浓度0.6 g/L、发酵温度30 ℃。果胶酶粗酶液、漆酶粗酶液酶解膨胀梗丝中果胶和木质素的最优条件分别为果胶酶粗酶液添加量240 U/g、酶解时间4.0 h、液质比1∶2、酶解温度30 ℃及漆酶粗酶液添加量2.0 U/g、酶解时间3.5 h、液质比1∶2、酶解温度30 ℃。膨胀梗丝酶解的最优粗酶液复配条件为果胶酶粗酶液添加量170 U/g、漆酶粗酶液添加量1.5 U/g、酶解时间3.0 h,酶解后其感官评吸得分较未酶解膨胀梗丝提高了8.2分,致香成分含量明显增加。Abstract: Pectinase-producing strain TS-1 and laccase-producing strain TS-2 were isolated from tobacco field soil and decayed tobacco stems. Morphological observation and molecular biological identification confirmed their taxonomic status as Aspergillus niger and Phanerodontia chrysosporium, respectively. Enzyme production conditions were optimized via single-factor experiments: for TS-1 pectinase production, the optimal parameters were tobacco stem powder concentration 50 g/L, fermentation temperature 30 ℃, and pH value 6.0; for TS-2 laccase production, the optimal conditions were tobacco stem powder concentration 50 g/L, (NH4)2SO4 concentration 0.6 g/L, and fermentation temperature 30 ℃. The hydrolysis conditions for pectin in expanded tobacco cut stems were optimized as crude pectinase dosage 240 U/g, hydrolysis time 4.0 h, liquid-to-solid ratio 1∶2, at 30 ℃; for lignin hydrolysis, the optimal conditions were crude laccase dosage 2.0 U/g, hydrolysis time 3.5 h, liquid-to-solid ratio 1∶2, at 30 ℃. The co-application of crude enzymes achieved optimal hydrolysis at pectinase dosage 170 U/g, laccase dosage 1.5 U/g, and hydrolysis time 3.0 h. Post-hydrolysis, the sensory evaluation score of expanded cut stems increased by 8.2 points compared to untreated samples, accompanied by a significant elevation in aroma constituent content.
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Key words:
- tobacco stem /
- pectinase /
- laccase /
- enzymatic hydrolysis process /
- expanded tobacco cut stem /
- sensory quality /
- aroma constituent
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