| 167 | 0 | 33 |
| 下载次数 | 被引频次 | 阅读次数 |
为了利用生物转化法合成绿色无毒的圆柚酮，从富含瓦伦西亚橘烯和圆柚酮的脐橙果皮中分离菌株，通过菌株的初筛、复筛选出能将瓦伦西亚橘烯转化成圆柚酮的特定菌株，利用16S rDNA测序对所筛菌株进行鉴定，并进行遗传稳定性测试和发酵条件优化。结果表明所筛菌株为山梨木糖假丝酵母；在吐温80添加量为4.00 g/L、瓦伦西亚橘烯添加量为30.00μL、pH为4.00，硫酸亚铁添加量为0.50 g/L，苹果酸添加量为30.00 g/L组成的发酵液下发酵48 h，山梨木糖假丝酵母利用瓦伦西亚橘烯生成圆柚酮的产量最高，为60.58mg/L。此外，该研究发现镁离子的存在对山梨木糖假丝酵母利用瓦伦西亚橘烯合成圆柚酮起抑制作用，亚铁离子起促进作用，为圆柚酮的工业化生产提供进一步的理论基础。
Abstract：To use biotransformation method to produce non-toxic nootkatone,strains were isolated from the navel orange peels rich in valencene and nootkatone. Through primary and secondary screening,strains capable of converting valencene into nootkatone were selected. The screened strains were identified by 16S rDNA sequencing and subjected to genetic stability tests and fermentation condition optimization. The results indicated that the most efficient strain was identified as Starmerella sorbosivorans. The optimal fermentation conditions were as follows :4.00 g/L Tween 80,30.00 μL valencene,pH 4.00,0.50 g/L ferrous sulfate,30.00 g/L malic acid and fermentation duration 48 h. Under the above conditions,the highest nootkatone yield reached 60.58 mg/L. Furthermore,the presence of magnesium ions was found to inhibit nootkatone synthesis by Starmerella sorbosivorans from valencene,while ferrous ions promoted it. These findings provide a foundation for the industrial production of nootkatone.
[1]Erdtman H,Hirose Y,Theander O. The chemistry of the natural order Cupressales. 46. The structure of nootkatone[J].Acta Chemica Scandinavica,1962,16:1311-1314.
[2]刘慧.酿酒酵母合成植物倍半萜类圆柚酮代谢途径的设计与构建[D].济南：山东大学，2019.
[3]Villafa?e E,Tolosa D,Bardón A,et al. Toxic effects of Citrus aurantium and C. Limon essential oils on Spodoptera frugiperda(Lepidoptera:Noctuidae)[J]. Natural Product Communications,2011,6(9):1389-1392.
[4]Castillo-Araiza C O,Palmerín-Carre?o D,Prado-Barragán A,et al. On the conceptual design of a partitioning technology for the bioconversion of(+)-valencene to(+)-nootkatone on whole cells:Experimentation and modelling[J]. Chemical Engineering and Processing:Process Intensification,2017,122:493-507.
[5]Ramachanderan R,Schaefer B.(+)-nootkatone,the flavor of grapefruit[J]. ChemTexts,2024,10(4):11.
[6]Belhassen E,Baldovini N,Brevard H,et al. Unravelling the scent of vetiver:Identification of character-impact compounds[J]. Chemistry&Biodiversity,2014,11(11):1821-1842.
[7]Liao J F,Zhao X Y. Recent research progress on the chemical constituents,pharmacology,and pharmacokinetics of alpinae oxyphyllae fructus[J]. Molecules,2024,29(16):3905.
[8]Fraatz M A,Riemer S J L,St?ber R,et al. A novel oxygenase from Pleurotus sapidus transforms valencene to nootkatone[J].Journal of Molecular Catalysis B:Enzymatic,2009,61(3-4):202-207.
[9]Beier C M,Sink S E,Hennon P E,et al. Twentieth-century warming and the dendroclimatology of declining yellow-cedar forests in southeastern Alaska[J]. Canadian Journal of Forest Research,2008,38(6):1319-1334.
[10]Liu T,Li W,Chen H F,et al. Systematic optimization of HPO-CPR to boost(+)-nootkatone synthesis in engineered Saccharomyces cerevisiae[J]. Journal of Agricultural and Food Chemistry,2022,70(49):15548-15559.
[11]王帅斌.高速逆流色谱分离益智中诺卡酮香味物质的研究[D].北京：北京工商大学，2009.
[12]Shaffer G W,Eschinasi E H,Purzycki K L,et al. Oxidations of valencene[J]. The Journal of Organic Chemistry,1975,40(15):2181-2185.
[13]Fraatz M A,Berger R G,Zorn H. Nootkatone:A biotechnological challenge[J]. Applied Microbiology and Biotechnology,2009,83(1):35-41.
[14]卢玥，杨易坤，郑若欣，等.产香酵母的筛选鉴定、生长特性及其混菌发酵果酒应用[J].应用与环境生物学报，2025,31(2):282-294.
[15]舒学香，周文，吴霞，等.猕猴桃果酒酿造专用酵母菌株的筛选[J].中国酿造，2021,40(8):99-104.
[16]郑裕国，孙丽慧，舒学香，等.山梨木糖假丝酵母及其在制备（s）-4-氯-3-羟基丁酸乙酯中的应用：201210594514.5[P]. 2012-12-31.
[17]Makhubela I M,Zawaira A,Brady D,et al. Multifactorial optimization enables the identification of a greener method to produce(+)-nootkatone[J]. Journal of Biotechnology,2024,393:41-48.
[18]Mukamolova G V,Kaprelyants A S,Young D I,et al. A bacterial cytokine[J]. Proceedings of the National Academy of Sciences,1998,95(15):8916-8921.
[19]袁麒，王旭苹，杨怀谷，等.不同pH条件下乳清蛋白-桑椹多酚复合营养粉的制备及其理化性质[J].现代食品科技，2023,39(5):164-174.
[20]张磊，罗泽华，杨明川，等.雪茄烟叶原料发酵微生物多样性及酶活变化研究[J].中国农业科技导报，2021,23(10):171-180.
[21]周德庆.微生物学教程（第4版）[M].北京：高等教育出版社，2020.
[22]梁安健，石沁兰，王金丽，等.产蛋白酶波茨坦短芽孢杆菌的鉴定及产酶条件优化[J].现代食品科技，2024,40(5):73-83.
[23]张晓瑞，刘晓晖，付博，等.烟草中淀粉降解菌的筛选、鉴定及发酵工艺优化[J].食品与机械，2021,37(2):34-41.
[24]李晓.瓦伦西亚橘烯生物转化生成圆柚酮的研究[D].武汉：华中农业大学，2019.
[25]Ivanov B,Asada K,Edwards G E. Analysis of donors of electrons to photosystem I and cyclic electron flow by redox kinetics of P700 in chloroplasts of isolated bundle sheath strands of maize[J]. Photosynthesis Research,2007,92(1):65-74.
[26]Johnston M. Feasting,fasting and fermenting. Glucose sensing in yeast and other cells[J]. Trends in Genetics,1999,15(1):29-33.
[27]李雪梅，康萌，万静之，等.酒精性肝病中的铁死亡机制研究进展[J].癌变·畸变·突变，2024,36(1):66-69,76.
[28]相启森，岳田利，袁亚宏，等.金属离子对褐黄孢链霉菌生长和纳他霉素生物合成的影响[J].西北农林科技大学学报：自然科学版，2010,38(1):209-215.
[29]魏琦.雄烯二酮11α羟化的球孢白僵菌菌株筛选及其转化工艺研究[D].武汉：华中农业大学，2007.
[30]蒋武成，萧克宇，徐文元.硫酸亚铁对某些病原菌的杀灭试验[J].湖南农学院学报，1991,17(2):197-200.
[31]向梦杰.合成谷蓝工程菌株的构建及全细胞催化[D].福州：福建师范大学，2021.
[32]田景芝.蜂蜜酒酵母菌株的筛选及发酵工艺的研究[D].福州：福建农林大学，2010.
[33]徐晓敏.毕赤酵母表面展示脂肪酶CALB全细胞催化合成肉桂醇酯[D].广州：华南理工大学，2018.
[34]Girhard M,Machida K,Itoh M,et al. Regioselective biooxidation of(+)-valencene by recombinant E. coli expressing CYP109B1 from Bacillus subtilis in a two-liquidphase system[J]. Microbial Cell Factories,2009,8:36.
基本信息:
DOI：10.19804/j.issn1006-2513.2025.11.007
中图分类号:TS264.3
引用信息:
[1]潘奕丞,刘晓旋,钱敏,等.产圆柚酮菌种筛选及发酵条件优化[J].中国食品添加剂,2025,36(11):48-58.DOI:10.19804/j.issn1006-2513.2025.11.007.
基金信息:
功能性内酯与手性香料绿色制备关键技术研发与示范（2022B0202050003）; 仲恺农业工程学院研究生创新基金（KJCX2024017）