类芽孢杆菌发酵原料浸提液提升再造烟叶品质
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摘要
为提升再造烟叶品质,利用烟草中分离的类芽孢杆菌制备静息细胞,对再造烟叶原料浸提液进行发酵处理;采用连续流动分析、气质联用仪(GC-MS)、热裂解-气相色谱质谱(Py-GC/MS)等方法,测定了发酵过程中浸提液中蛋白质、还原糖、氨基酸含量变化;分析了其主要挥发性香味成分;并利用发酵浸提液浓缩涂布制备再造烟叶样品,进行热裂解产物分析和感官评价。结果表明:①发酵浸提液和对照组中的蛋白质、还原糖含量在5 h内持续减少,而氨基酸含量先增高后降低,且发酵浸提液的蛋白质、还原糖含量低于对照,氨基酸含量高于对照;②发酵液中香味物质种类和总量较对照组均有所增加,其中糠醛、糠酮等致香组分增幅明显;③对比再造烟叶裂解产物,发现发酵浸提液浓缩后涂布样品裂解产生的γ-丁内酯、2,3-二氢-5-甲基呋喃、糠醛等典型烟气致香组分提高;④发酵液浓缩后涂布的再造烟叶样品感官品质较对照样品明显提升。利用类芽孢杆菌发酵烟草浸提液,对于提升再造烟叶的感官品质具有促进作用。
In order to improve quality of reconstituted tobacco, resting cells of a bacterium Paenibacillus sp. isolated from tobacco leaves were used for the fermentation of tobacco extract during production of reconstituted tobacco. Different analytical methods such as continuous ?owing analysis, gas chromatography mass spectrometry(GC/MS) and pyrolysis gas chromatography mass spectrometry(PY-GC/MS) were applied.Contents of protein, reducing sugar and amino acid in tobacco extract was monitored, and its ?avor ingredients were detected. Reconstituted tobacco was prepared by coating concentrated tobacco extract fermentation broth. Pyrolysis product analysis and sensory evaluation of final reconstituted tobacco was also carried out to assess effect of bacterium fermentation. Results showed that: 1) In 5 hours, contents of protein,reducing sugar in tobacco extract reduced while amino acid increased ?rst and then decreased. Nevertheless, the fermentation tobacco extract(adding Paenibacillus sp.) featured lower contents of protein and reducing sugar but higher contents of amino acid than control tobacco extract(without adding Paenibacillus sp.). 2) Compared with control tobacco extract, the species and quantities of aroma compounds in fermented tobacco extract were higher, among which furfural aldehydes and keltones were increased. 3) Aroma compounds of pyrolysis products, such as4-Butanolide, 2,3-Dihydro-5-methyl-Furan, Furfural were increased in ?nal reconstituted tobacco coated with fermentation tobacco extract. 4)Compared with control reconstituted tobacco, sensory quality of ?nal reconstituted tobacco coated with fermentation tobacco extract were also improved. Thus, tobacco extract fermentation by Paenibacillus sp. during production process could increase ?nal quality of reconstituted tobacco.
In order to improve quality of reconstituted tobacco, resting cells of a bacterium Paenibacillus sp. isolated from tobacco leaves were used for the fermentation of tobacco extract during production of reconstituted tobacco. Different analytical methods such as continuous ?owing analysis, gas chromatography mass spectrometry(GC/MS) and pyrolysis gas chromatography mass spectrometry(PY-GC/MS) were applied.Contents of protein, reducing sugar and amino acid in tobacco extract was monitored, and its ?avor ingredients were detected. Reconstituted tobacco was prepared by coating concentrated tobacco extract fermentation broth. Pyrolysis product analysis and sensory evaluation of final reconstituted tobacco was also carried out to assess effect of bacterium fermentation. Results showed that: 1) In 5 hours, contents of protein,reducing sugar in tobacco extract reduced while amino acid increased ?rst and then decreased. Nevertheless, the fermentation tobacco extract(adding Paenibacillus sp.) featured lower contents of protein and reducing sugar but higher contents of amino acid than control tobacco extract(without adding Paenibacillus sp.). 2) Compared with control tobacco extract, the species and quantities of aroma compounds in fermented tobacco extract were higher, among which furfural aldehydes and keltones were increased. 3) Aroma compounds of pyrolysis products, such as4-Butanolide, 2,3-Dihydro-5-methyl-Furan, Furfural were increased in ?nal reconstituted tobacco coated with fermentation tobacco extract. 4)Compared with control reconstituted tobacco, sensory quality of ?nal reconstituted tobacco coated with fermentation tobacco extract were also improved. Thus, tobacco extract fermentation by Paenibacillus sp. during production process could increase ?nal quality of reconstituted tobacco.
引文
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[11]GB/T8314-87茶游离氨基酸总量测定[S].GB/T8314-87 Determination of total free amino acids in tea[S].
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[18]李丹,黄龙,朱巍,等.反相高效液相色谱法测定烟叶中的游离氨基酸[J].烟草科技,2003(2):20-24.LI Dan,HUANG Long,ZHU Wei,et al.Determination of free amino acids in tobacco leaves by Reversed Phase High Performance Liquid Chromatography[J].Tobacco Science&Techonology,,2003(2):20-24.
[2]唐杰斌,赵传山,韩文佳.造纸法再造烟叶生产工艺的改进[J].烟草科技,2009(7):15-17.TANG Jiebin,ZHAO Chuanshan,HAN Wenjia.Improvement of production process of paper making reconstituted tobacco[J].Tobacco Science&Technology,2009(7):15-17.
[3]李丹,刘熙.生物技术应用于造纸法再造烟叶生产的研究进展[J].现代食品科技,2013,29(6):1463-1466.LI Dan,LIU Xi.Research progress of biological technology applied to paper making reconstituted tobacco production[J].Modern Food Science&Technology,2013,29(6):1463-1466.
[4]朱国成.酶处理技术在造纸法烟草再造烟叶中的应用研究[D].北京:中国农业大学,2006.ZHU Guocheng.Application of enzyme treatment technology in tobacco reconstituted tobacco by papermaking process[D].Beijing:China Agricultural University,2006.
[5]吴亦集,沈光林,陶红,等.造纸法再造烟叶原料的加酶萃取[J].烟草科技,2011(7):33-36.WU Yiji,SHEN Guanglin,TAO Hong,et al.Enzymatic extraction of raw materials in papermaking process[J].Tobacco Science&Technology,2011(7):33-36.
[6]张勃,贾玉红,端李祥,等.微生物发酵烟梗水提物的制备及其在再造烟叶中的应用[J].河南农业科学,2012,41(3):56-60.ZHANG Bo,JIA Yuhong,DUAN Lixiang,et al.Study on preparation of fermented tobacco stem extracts and application to reconstituted tobacco.[J].Journal of Henan Agricultural Science,2012,41(3),56-60.
[7]梁艳玲.产纤维素酶细菌的分离、鉴定及地类芽孢杆菌ME27-1产酶条件优化[D].南宁:广西大学,2014.LIAN Yanling.Isolation and identification of Cellulase Producing Bacteria and optimization of conditions for production of Bacillus subtilis ME27-1[D].Nanning:Guangxi University,2014.
[8]刘飞,夏振远,白江兰,等.土壤中降解烟碱细菌多样性研究[J].中国烟草学报,2012,18(2):58-64.LIU Fei,XIA Zhenyuan,BAI Jianglan,et al.Study on bacterial diversity of nicotine degradation in soil[J].Acta Tabacaria Sinica,2012,18(2):58-64.
[9]YE Jianbin,ZHANG Zhan,YAN Ji,et al.Degradation of phytosterols in tobacco waste extract by a novel Paenibacillus sp[J].Biotechnology&Applied Biochemistry,2017,64(6):843-850.
[10]王文领,李华雨.再造烟叶质量稳定技术[M].北京:化学工业出版社,2016.WANG Wenling,LI Huayu.Stable quality of reconstituted tobacco technology[M].Beijjing:Chemical Industry Press,2016.
[11]GB/T8314-87茶游离氨基酸总量测定[S].GB/T8314-87 Determination of total free amino acids in tea[S].
[12]YC/T159-2002烟草及烟草制品水溶性糖的测定连续流动法[S].YC/T159-2002 Tobacco and tobacco products Determination of water soluble sugars by continuous flow method[S]
[13]杨静,白冰,王宁,等.考马斯亮蓝法对烟草薄片涂布液中蛋白质含量的测定[J].湖北农业科学,2017,56(5):946-947,950.YANG Jing,BAI Bing,WANG Ning,et al.Determination of protein content in reconstituted tobacco coating liquid by coomassie brilliant blue method[J].Hubei Agricultural Sciences,2017,56(5):946-947,950.
[14]YC/T498-2014再造烟叶(造纸法)感官评价方法[S].YC/T498-2014 The evaluation methods of sensory quality for reconstituted tobacco(paper process)[S].
[15]谢剑平.烟草香原料[M].北京:化学工业出版社,2009.7.XIE Jianping.Tobacco flavoring material[M].Beijing:Chemical Industry Press,2009.
[16]舒明,樊虎,刘金莉,等.基于高通量测序的废次烟末水提液中细菌群落分析[J].烟草科技,2016,49(4);1-7.SHU Ming,FAN Hu,LIU Jinli,et.al.High throughput bacterial analysis on aqueous extract of waste tobacco scrap[J].Tobacco Science&Techonology,2016,49(4):1-7.
[17]马戎,鲍峰伟,冯文宁,等.液相色谱串联质谱法测定烟草中的游离氨基酸[J].中国测试,2013,39(2):34-37.MA Rong,BAO Fengwei,FENG Wenning,et al.Determination of free amino acids in tobacco by liquid chromatography tandem mass spectrometry[J].China Measurement&Testing Technology,2013,39(2):34-37.
[18]李丹,黄龙,朱巍,等.反相高效液相色谱法测定烟叶中的游离氨基酸[J].烟草科技,2003(2):20-24.LI Dan,HUANG Long,ZHU Wei,et al.Determination of free amino acids in tobacco leaves by Reversed Phase High Performance Liquid Chromatography[J].Tobacco Science&Techonology,,2003(2):20-24.