三菌混合发酵结合美拉德反应对烟末提取液的提质研究
首发时间:2017-02-06
摘要:烟末提取液是烟草薄片重要的香气来源,传统提取法所得提取液香气含量低,残渣中有害前体物质难以降解。本文的研究目的是筛选产纤维素酶、果胶酶和漆酶的优良菌株,将其作为提质改良剂进行混合发酵并结合美拉德反应来改善烟末提取液的品质。通过平板分离法从五种不同产地的醇化烟叶中筛选出产纤维素酶、果胶酶和漆酶的优势菌株各一株,粗酶液酶活分别可达199.90 U/mL, 136.30 U/mL和7.86 U/L。采用16S rDNA序列分析结合常规鉴定方法对三株菌进行鉴定,均属于枯草芽孢杆菌。三株菌在pH 6.0, 35℃下按2 %接种量于NA 培养基中先分别培养12 h,再按体积比1∶2∶1的比例混合培养至24 h所得粗酶液各酶活均较高。将混合菌液接入烟末提取液发酵后,烟末提取率提高3.63%,残渣中纤维素、果胶、木质素分别减少17.87%、52.39%和23.90%;提取液中还原糖增加20.18%。烟末提取液发酵后进行美拉德反应,愉悦香气成分增加了150.05%;不愉悦香气成分降低了70.67%,此技术对烟末提取液提质效果显著。
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Study on quality improvement of tobacco powder extract by three - bacteria mixed fermentation and Maillard reaction
Abstract:The extraction of tobacco powder is an important aroma source for tobacco sheets. The aroma content of extracts from traditional extraction is low, and the harmful precursors in the residue are difficult to degrade. The purpose of this study is to screen the strains secreting cellulase, pectinase and laccase for mixed fermentation and Maillard reaction to improve the quality of tobacco powder extract. Using the plate separation method, three strains were screened from five types of tobacco grown in different places. The crude enzymatic activity of cellulase, pectinase and laccase was 199.90 U/mL, 136.30 U/mL and 7.86 U/L, respectively. They were all identified as Bacillus subtilis using 16S rDNA sequence analysis combined with routine identification methods. The activity of the crude enzyme was higher on condition that three strains were incubated in NA with 2% inoculum at pH 6.0 and 35℃ for 12 h respectively, then mixed with 1: 2: 1 ratio for 24 h. After fermentation, the extraction yield of tobacco powder was increased by 3.63%, and the content of cellulose, pectin and lignin in residue was decreased by 17.87%, 52.39% and 23.90%, respectively. The reducing sugar was increased by 20.18% in extracting solution. Combined with Maillard reaction, the desirable aroma components were increased by 150.05% and the undesirable components were decreased by 70.67%, which proved significant improvement to the extraction of tobacco powder.
Keywords: tobacco powder extraction mixed fermentation Maillard reaction GC-MS
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