烟草科技2026，Vol.59Issue(3)：44-53,10.DOI:10.16135/j.issn1002-0861.2025.0461
产香β-胡萝卜素双加氧酶挖掘与功能验证及酶学性质分析
Discovery,functional verification and enzymatic properties of β-carotene dioxygenases with aroma-producing capability
摘要
Abstract
To screen carotenoid cleavage dioxygenases(CCDs)specifically capable of degrading β-carotene to produce aroma substances,a bioinformatic search based on structural similarity was employed with Petunia hybrida PhCCD1 as the reference sequence,10 novel CCDs candidates were identified from the public metaproteomic database MGnify.Heterologous expression in Escherichia coli confirmed successful production of 9 out of 11 recombinant CCD proteins,with PhCCD1 and CCD-11 exhibiting soluble expression.In vitro enzymatic results showed that both PhCCD1 and CCD-11 displayed strong degradation activity toward β-carotene and zeaxanthin.Gas chromatography-mass spectrometry(GC-MS)analysis revealed that these enzymes catalyzed the cleavage production of β-ionone from β-carotene,with yields reaching 637.35 and 523.92 μg/mL,respectively.In addition,both enzymes demonstrated lycopene degradation capability.Further characterization showed that CCD-11 demonstrated optimal activity at 50℃with a pH value of 5.0,and its enzymatic activity was significantly enhanced by Fe2+and Tween-80.The Michaelis constant(Km)of CCD-11 was 4.60 mol/L,and the maximum enzymatic reaction rate(Vmax)was 3.47 μmol/(min·mg).This study demonstrates that the novel CCD-11 possesses the ability to produce the aroma substance β-ionone,providing a research foundation for its application in tobacco aroma enhancement.关键词
β-胡萝卜素/类胡萝卜素裂解双加氧酶/微生物/气相色谱-质谱/β-紫罗兰酮Key words
β-carotene/Carotenoid cleavage dioxygenase/Microorganism/Gas chromatography-mass spectrometry/β-ionone分类
轻工纺织引用本文复制引用
董盼盼,龙腾,钟家威,谢金栋,何伟,林志平,陈千思,许亚龙,李菁菁..产香β-胡萝卜素双加氧酶挖掘与功能验证及酶学性质分析[J].烟草科技,2026,59(3):44-53,10.基金项目
福建中烟工业有限责任公司科技项目"烟草质体色素降解菌筛选构建及功能酶制剂开发"(FJZYKJJH2023ZD006) （FJZYKJJH2023ZD006）
中国烟草总公司重大科技项目"烟草典型共栖微生物资源鉴定、特征解析与开发利用"[110202201004(JY-04)]. （JY-04）
