柠檬罗勒产香细菌发酵条件优化及卷烟加香应用
Optimization of fermentation conditions of aroma-producing endophytic bacteria in Ocimum × citriodorum and its application in cigarette flavoring
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摘要: 【目的】 挖掘柠檬罗勒(Ocimum×citriodorum)内生细菌的产香潜力。【方法】 采用传统分离培养结合形态学与分子生物学方法,从柠檬罗勒根、茎、叶中筛选鉴定优势产香菌株,通过单因素试验与响应面法优化发酵条件,利用同时蒸馏萃取(SDE)法提取挥发油并进行卷烟加香评价。【结果】 共分离获得38株内生细菌,感官评价确定菌株J-3-6的产香品质最佳,经鉴定为菠萝泛菌(Pantoea ananatis),呈现清甜茉莉花香气,主要产香物质为吲哚,最优发酵条件为乳糖质量浓度10.2 g/L、酵母浸粉质量浓度50.7 g/L、装液量259 mL、接种量7%、初始pH 值7、发酵温度26 ℃、摇床转速180 r/min和发酵时间48 h,在此条件下,吲哚产量是初始产量的5.53倍,共发酵体系鉴定出45种挥发性有机化合物,以醇类、酯类和杂环类为主,香气特征在清新果香的基础上融合了复杂花香与木质香调,卷烟加香评价表明最适宜添加量为4 μg/支。【结论】 菠萝泛菌J-3-6经响应面法发酵条件优化可显著提升吲哚产量,其代谢产物挥发油可醇和烟香,降低卷烟杂气与刺激性。Abstract: 【Objective】 This study aims to explore the aroma-producing potential of endophytic bacteria in Ocimum × citriodorum. 【Methods】 Traditional isolation and culture method combined with morphological and molecular biological method were used to screen and identify the dominant aroma-producing strains from its roots, stems and leaves; then, the fermentation conditions were optimized by the single factor test and response surface method; and volatile oil was extracted by simultaneous distillation extraction (SDE) and subjected to cigarette flavoring evaluation. 【Results】 A total of 38 strains of endophytic bacteria were isolated. Sensory evaluation confirmed that strain J-3-6 had the best aroma quality, which was identified as Pantoea ananatis exhibiting a sweet jasmine aroma with indole as the main aroma-producing substance. The optimal fermentation conditions were as follows: lactose mass concentration of 10.2 g/L, yeast extract powder mass concentration of 50.7 g/L, liquid volume of 259 mL, inoculation amount of 7%, initial pH value of 7, fermentation temperature of 26 ℃, shaker rotation speed of 180 r/min, and fermentation time of 48 h. Under these conditions, the indole yield was 5.53 times the initial yield. A total of 45 volatile organic compounds were identified in the fermentation system, mainly composed of alcohols, esters and heterocyclic compounds. The aroma characteristics were based on a fresh fruity aroma, integrated with complex floral and woody notes. Cigarette flavoring evaluation showed that the most suitable addition level was 4 μg/cigarette. 【Conclusion】 Optimization of the fermentation conditions for Pantoea ananatis J-3-6 by the response surface method significantly enhances indole production, and its metabolites extracted as volatile oils can effectively harmonize tobacco aroma and reduce the offensive odor and irritation of cigarettes.
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