烟草NtDHQ/SDH8基因突变对卷烟主流烟气中苯酚释放量的影响

TS413; 为明确莽草酸途径的重要双功能酶3-脱氢奎尼酸脱水酶/莽草酸脱氢酶(DHQ/SDH)突变对酪氨酸合成和卷烟主流烟气中苯酚等酚类化合物释放量的影响,使用TILLING技术在烟草EMS突变体库中筛选出14个ntdhq/sdh8突变体,分析其突变位点和突变导致的蛋白质功能变异,测定烤后烟叶的酪氨酸、脯氨酸、天冬酰胺和总蛋白质的含量(质量分数),以及卷烟主流烟气中苯酚、邻-苯二酚、对-苯二酚、间-苯二酚、间/对-甲酚和邻-甲酚的释放量.结果显示,突变体NtDSM03、NtDSM12和NtDSM13中NtDHQ/SDH8双功能酶的DHQ结构域氨基酸发生变异,导致蛋白三级结构以及酶活性发生变化...

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Published in烟草科技 Vol. 57; no. 12; pp. 42 - 48
Main Authors 李锋, 刘楠, 樊武广, 姜兴益, 王燃, 金立锋, 李正风, 王大伟
Format Journal Article
LanguageChinese
Published 中国烟草总公司郑州烟草研究院,郑州高新技术产业开发区枫杨街2号 450001%国家烟草质量监督检验中心,郑州高新技术产业开发区翠竹街6号 450001%河南省烟草公司许昌市公司,河南省许昌市魏都区湖滨路43号 461099%河南农业大学生命科学学院,郑州市郑东新区平安大道218号 450046%云南中烟工业有限责任公司技术中心,昆明市红锦路367号 650231%中国农业科学院烟草研究所 烟草行业烟草基因资源利用重点实验室,山东省青岛市崂山区科苑经四路11号 266101 01.12.2024
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ISSN1002-0861
DOI10.16135/j.issn1002-0861.2024.0287

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Abstract TS413; 为明确莽草酸途径的重要双功能酶3-脱氢奎尼酸脱水酶/莽草酸脱氢酶(DHQ/SDH)突变对酪氨酸合成和卷烟主流烟气中苯酚等酚类化合物释放量的影响,使用TILLING技术在烟草EMS突变体库中筛选出14个ntdhq/sdh8突变体,分析其突变位点和突变导致的蛋白质功能变异,测定烤后烟叶的酪氨酸、脯氨酸、天冬酰胺和总蛋白质的含量(质量分数),以及卷烟主流烟气中苯酚、邻-苯二酚、对-苯二酚、间-苯二酚、间/对-甲酚和邻-甲酚的释放量.结果显示,突变体NtDSM03、NtDSM12和NtDSM13中NtDHQ/SDH8双功能酶的DHQ结构域氨基酸发生变异,导致蛋白三级结构以及酶活性发生变化.与野生型相比,突变体NtDSM03和NtDSM12烤后烟叶的酪氨酸质量分数分别降低76.78%和81.71%,卷烟主流烟气中苯酚释放量分别降低24.09%和48.99%,而且降幅均达到极显著水平(P<0.01).突变体NtDSM12烤后烟叶的酪氨酸、总蛋白质质量分数以及卷烟主流烟气中苯酚、邻-苯二酚、间/对-甲酚和邻-甲酚等4种酚类化合物的释放量均极显著下降(P<0.01).
AbstractList TS413; 为明确莽草酸途径的重要双功能酶3-脱氢奎尼酸脱水酶/莽草酸脱氢酶(DHQ/SDH)突变对酪氨酸合成和卷烟主流烟气中苯酚等酚类化合物释放量的影响,使用TILLING技术在烟草EMS突变体库中筛选出14个ntdhq/sdh8突变体,分析其突变位点和突变导致的蛋白质功能变异,测定烤后烟叶的酪氨酸、脯氨酸、天冬酰胺和总蛋白质的含量(质量分数),以及卷烟主流烟气中苯酚、邻-苯二酚、对-苯二酚、间-苯二酚、间/对-甲酚和邻-甲酚的释放量.结果显示,突变体NtDSM03、NtDSM12和NtDSM13中NtDHQ/SDH8双功能酶的DHQ结构域氨基酸发生变异,导致蛋白三级结构以及酶活性发生变化.与野生型相比,突变体NtDSM03和NtDSM12烤后烟叶的酪氨酸质量分数分别降低76.78%和81.71%,卷烟主流烟气中苯酚释放量分别降低24.09%和48.99%,而且降幅均达到极显著水平(P<0.01).突变体NtDSM12烤后烟叶的酪氨酸、总蛋白质质量分数以及卷烟主流烟气中苯酚、邻-苯二酚、间/对-甲酚和邻-甲酚等4种酚类化合物的释放量均极显著下降(P<0.01).
Abstract_FL To determine the effects of mutations in 3-dehydroquinic acid dehydrase/shikimate dehydrogenase(DHQ/SDH),an important bifunctional enzyme in the shikimate pathway,on tyrosine synthesis and the release of phenol and other phenolic compounds in mainstream cigarette smoke,fourteen ntdhq/sdh8 mutants were screened and identified by TILLING technology in the EMS mutagenesis population of tobacco(Nicotiana tabacum L.).The mutation sites and the functional variation of the proteins caused by the mutation were analyzed,and the mass fractions of tyrosine,proline,asparagine,and total protein,as well as the release of phenol,o-dihydroxybenzene,p-dihydroxybenzene,m-dihydroxybenzene,m/p-cresol,and o-cresol in mainstream cigarette smoke were determined.The results showed that the tertiary structure of the DHQ functional domain in NtDHQ/SDH8 protein was altered by amino acid variation,which affected the enzymatic activity of the three ntdhq/sdh8 mutants.Compared with the wild-type control,the tyrosine mass fraction in NtDSM03 and NtDSM12 mutants decreased by 76.78%and 81.71%,respectively;and the phenol release from mainstream cigarette smoke of the two mutants decreased by 24.09%and 48.99%,respectively;all of which showed extremely significant differences(P<0.01).In particular,the mass fractions of total protein,tyrosine,and phenolic compounds such as phenol,catechol,m/p-cresol,and o-cresol in the mainstream cigarette smoke of the NtDSM12 mutant decreased extremely significantly compared to the wild-type control(P<0.01).
Author 李锋
刘楠
金立锋
王燃
樊武广
李正风
姜兴益
王大伟
AuthorAffiliation 中国烟草总公司郑州烟草研究院,郑州高新技术产业开发区枫杨街2号 450001%国家烟草质量监督检验中心,郑州高新技术产业开发区翠竹街6号 450001%河南省烟草公司许昌市公司,河南省许昌市魏都区湖滨路43号 461099%河南农业大学生命科学学院,郑州市郑东新区平安大道218号 450046%云南中烟工业有限责任公司技术中心,昆明市红锦路367号 650231%中国农业科学院烟草研究所 烟草行业烟草基因资源利用重点实验室,山东省青岛市崂山区科苑经四路11号 266101
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Author_FL WANG Dawei
LI Feng
FAN Wuguang
LI Zhengfeng
LIU Nan
JIANG Xingyi
WANG Ran
JIN Lifeng
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DocumentTitle_FL Effects of NtDHQ/SDH8 mutations on phenol release in mainstream cigarette smoke
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Keywords Tyrosine
烟草突变体
Phenol
3-Dehydroquinate dehydratase/shikimate dehydrogenase
酪氨酸
Mainstream cigarette smoke
3-脱氢奎尼酸脱水酶/莽草酸脱氢酶
酚类化合物
苯酚
Tobacco mutant
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Publisher 中国烟草总公司郑州烟草研究院,郑州高新技术产业开发区枫杨街2号 450001%国家烟草质量监督检验中心,郑州高新技术产业开发区翠竹街6号 450001%河南省烟草公司许昌市公司,河南省许昌市魏都区湖滨路43号 461099%河南农业大学生命科学学院,郑州市郑东新区平安大道218号 450046%云南中烟工业有限责任公司技术中心,昆明市红锦路367号 650231%中国农业科学院烟草研究所 烟草行业烟草基因资源利用重点实验室,山东省青岛市崂山区科苑经四路11号 266101
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Snippet TS413; 为明确莽草酸途径的重要双功能酶3-脱氢奎尼酸脱水酶/莽草酸脱氢酶(DHQ/SDH)突变对酪氨酸合成和卷烟主流烟气中苯酚等酚类化合物释放量的影响,使用TILLING技术在烟...
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Title 烟草NtDHQ/SDH8基因突变对卷烟主流烟气中苯酚释放量的影响
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