CN116376950A - 一种核酸构建体、细胞和四氢姜黄素的制备方法及应用 - Google Patents
一种核酸构建体、细胞和四氢姜黄素的制备方法及应用 Download PDFInfo
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Abstract
一种核酸构建体、细胞和四氢姜黄素的制备方法及应用,涉及生物工程技术领域;转化四氢姜黄素用核酸构建体,包括编码姜黄素还原酶的基因和编码用于还原辅酶的脱氢酶的基因。本发明的一种转化四氢姜黄素用核酸构建体,可在宿主细胞中表达姜黄素还原酶和用于还原辅酶的脱氢酶,姜黄素还原酶可将姜黄素转化为四氢姜黄素,同时还原型辅酶参与上述转化进程,并生成氧化型辅酶,所述用于还原辅酶的脱氢酶只需摄取经济易得的葡萄糖、葡萄糖酸、甲酸盐或异丙醇等反应底物即可使氧化型辅酶进行还原,从而构建辅酶循环系统,大幅提升四氢姜黄素的转化率,具有原材料可再生、对环境友好、反应条件温和、选择性强、安全性高的特点。
Description
技术领域
本发明属于生物工程技术领域,具体涉及一种核酸构建体、细胞和四氢姜黄素的制备方法及应用。
背景技术
姜黄素(Curcumin,1,7-二(4-羟基-3-甲氧基)苯基-1,6-庚二烯-3,5-二酮),普遍存在姜科、天南星科植物的块根或根茎中,是一种具有抗炎、抗癌、抗氧化、抗糖尿病、抗艾滋和抗阿尔兹海默症功能的天然多酚类化合物,来源易得。
由于姜黄素存在水溶性低、生物利用度低、易降解、稳定性差的问题,阻碍了其在临床上的广泛应用,因此,有必要研究姜黄素的衍生物,即保留姜黄素原有功能,又可适用于不同的应用场景。
四氢姜黄素(Tetrahydrocurcumin,THC),作为姜黄素的众多衍生物之一,可通过姜黄素氢化获得,同样具有抗氧化、抗炎、抗肿瘤、抗动脉粥样硬化和增强胰岛细胞功能等药理活性。1978年,Holder证实了四氢姜黄素与姜黄素的有类似药理作用。与姜黄素相比,四氢姜黄素在抗高血脂、抗糖尿病及抗炎方面的作用优于姜黄素,在抗氧化方面的作用等同于姜黄素,并可在较低的浓度下发挥作用,是具有广阔应用前景的姜黄素药物替代物。
目前,四氢姜黄素可通过化学合成法或生物转化法制得,通过化学合成法氢化姜黄素生成四氢姜黄素的方法危险且难以控制;利用生物转化法转化四氢姜黄素,具有原材料可再生、对环境友好、反应条件温和、选择性强、安全性高的特点,其可通过姜黄素还原酶催化姜黄素生成四氢姜黄素,但产量仅为毫克级别,该过程在还原型辅酶参与下可达到克级别,但是还原型辅酶价格昂贵、保存条件苛刻。
发明内容
为了克服现有技术的不足,本发明的目的之一在于提供一种转化四氢姜黄素用核酸构建体,用于辅酶的再生,构建辅酶循环系统,从而大幅提升催化生成四氢姜黄素的产率。
本发明的目的之二在于提供一种重组细胞。
本发明的目的之三在于提供一种转化四氢姜黄素用核酸构建体的应用。
本发明的目的之四在于提供一种四氢姜黄素的制备方法,反应条件温和、安全性高,且具有较高的成产效率。
本发明的目的之一采用如下技术方案实现:
一种转化四氢姜黄素用核酸构建体,包括编码姜黄素还原酶的基因和编码用于还原辅酶的脱氢酶的基因。
进一步地,所述姜黄素还原酶包括如下蛋白质(a)或(b):
(a)由氨基酸序列如SEQ ID NO:6所示的蛋白质;
(b)在(a)限定的氨基酸序列中经过取代、缺失或添加一种或几个氨基酸且具有姜黄素还原酶活性的由(a)衍生的具有至少95%序列同一性的同源蛋白质。
进一步地,所述姜黄素还原酶包括curA还原酶、yncB还原酶、yfeF还原酶、ybjS还原酶和ygfF还原酶中至少一种。
进一步地,编码所述yncB还原酶基因的核苷酸序列如SEQ ID NO:1所示。
进一步地,所述辅酶为烟酰胺腺嘌吟二核苷酸(NAD)和/或烟酰胺腺嘌吟二核苷磷酸(NADP)。
进一步地,所述还原辅酶的脱氢酶包括6-磷酸葡萄糖酸脱氢酶、葡萄糖脱氢酶、醇脱氢酶和甲酸脱氢酶中的至少一种。
进一步地,所述6-磷酸葡萄糖酸脱氢酶的氨基酸序列如SEQ ID NO:7所示;
所述葡萄糖脱氢酶的氨基酸序列如SEQ ID NO:8所示;
所述醇脱氢酶的氨基酸序列如SEQ ID NO:9所示;
所述甲酸脱氢酶的氨基酸序列如SEQ ID NO:10所示。
进一步地,编码6-磷酸葡萄糖酸脱氢酶的基因的核苷酸序列如SEQ ID NO:2所示;
编码葡萄糖脱氢酶的基因的核苷酸序列如SEQ ID NO:3所示;
编码醇脱氢酶的基因的核苷酸序列如SEQ ID NO:4所示;
编码甲酸脱氢酶的基因的核苷酸序列如SEQ ID NO:5所示。
本发明的目的之二采用如下技术方案实现:
一种重组细胞,所述重组细胞内所述的转化四氢姜黄素用核酸构建体。
本发明的目的之三采用如下技术方案实现:
一种转化四氢姜黄素用核酸构建体的应用,所述的一种转化四氢姜黄素用核酸构建体在姜黄素转化为四氢姜黄素的生产中的应用。
本发明的目的之四采用如下技术方案实现:
一种四氢姜黄素的制备方法,包括以下步骤:
S1,将所述的转化四氢姜黄素用核酸构建体重组到宿主细胞的表达载体中,然后对重组后的工程菌培养并诱导表达,得到催化酶液;
S2,将所述催化酶液与姜黄素和脱氢酶的反应底物混合并进行全细胞催化,生成四氢姜黄素。
相比现有技术,本发明的有益效果在于:
本发明的一种转化四氢姜黄素用核酸构建体,可在宿主细胞中表达姜黄素还原酶和用于还原辅酶的脱氢酶,姜黄素还原酶可将姜黄素转化为四氢姜黄素,同时还原型辅酶参与上述转化进程,并生成氧化型辅酶,所述用于还原辅酶的脱氢酶只需摄取经济易得的葡萄糖、葡萄糖酸、甲酸盐或异丙醇等反应底物即可使氧化型辅酶进行还原,从而构建辅酶循环系统,大幅提升四氢姜黄素的转化率,具有原材料可再生、对环境友好、反应条件温和、选择性强、安全性高的特点。
本发明的一种四氢姜黄素的制备方法,反应条件温和、安全性高,且具有较高的成产效率。
附图说明
图1是本发明的实施例2中pESC-HIS-yncB-6PGDH构建体的质粒谱图。
图2是本发明的实施例3中pET-28a-yncB-6PGDH构建体的质粒谱图。
图3是本发明的实施例3中姜黄素在反应前的外观图。
图4是本发明的实施例3中姜黄素在反应后的外观图。
图5是本发明的实施例4的全细胞催化产物的HPLC结果图。
图6是本发明的实施例5的全细胞催化产物的HPLC结果图。
图7是四氢姜黄素标准品的HPLC检测结果图。
具体实施方式
下面,结合具体实施方式,对本发明做进一步描述,需要说明的是,在不相冲突的前提下,以下描述的各实施例之间或各技术特征之间可以任意组合形成新的实施例。
一种转化四氢姜黄素用核酸构建体,包括编码姜黄素还原酶的基因和编码用于还原辅酶的脱氢酶的基因。
进一步地,所述姜黄素还原酶包括如下蛋白质(a)或(b):
(a)由氨基酸序列如SEQ ID NO:6所示的蛋白质;
(b)在(a)限定的氨基酸序列中经过取代、缺失或添加一种或几个氨基酸且具有姜黄素还原酶活性的由(a)衍生的具有至少95%序列同一性的同源蛋白质。
作为优选,所述姜黄素还原酶包括酶分类号EC:1.3.1中的酶,进一步地,所述姜黄素还原酶包括curA还原酶、yncB还原酶、yfeF还原酶、ybjS还原酶和ygfF还原酶中至少一种或其同源物。
如本文所用的,术语“序列同一性”是指两个或更多个氨基酸序列之间的关系。当一个序列中的一个位置被比较序列的对应位置中的相同氨基酸残基占据时,则这些序列被称为在该位置处是“同一的”。“序列同一性”百分比是通过以下方式计算:测定在两个序列中出现相同氨基酸残基的位置的数量而得出“同一的”位置的数量。然后,将“同一的”位置的数量除以比较窗口中位置的总数量,再乘以100,得出“序列同一性”的百分比。通过在比较窗口中比较两个最佳比对序列来确定“序列同一性”的百分比。为了最佳比对序列以进行比较,比较窗口中多肽序列的一部分可以包含称为空位的添加或缺失,而参考序列保持不变。最佳比对是这样的比对,其即使有空位也能在参考序列和比较序列之间产生最大可能的“同一”位置数量。可以使用已知方法计算编码序列之间的序列同一性水平。序列同一性可以使用用于测定序列同一性的可公开获得的基于计算机的方法来计算,包括BLASTP、BLASTN和FASTA(Atschul et al.,J.Molec.Biol.,215:403 410,(1990))、可由NCBI获得的BLASTX程序以及可由Genetics Computer Group(Madison WI)获得的Gap程序。使用Gap程序获得序列同一性的水平,其中氨基酸序列比较的Gap罚分是50,Gap长度罚分是3。
进一步地,编码所述yncB还原酶基因的核苷酸序列如SEQ ID NO:1所示。
进一步地,所述辅酶为烟酰胺腺嘌吟二核苷酸和/或烟酰胺腺嘌吟二核苷磷酸。
进一步地,所述还原辅酶的脱氢酶包括6-磷酸葡萄糖酸脱氢酶、葡萄糖脱氢酶、醇脱氢酶和甲酸脱氢酶中的至少一种。
进一步地,所述6-磷酸葡萄糖酸脱氢酶的氨基酸序列如SEQ ID NO:7所示;
所述葡萄糖脱氢酶的氨基酸序列如SEQ ID NO:8所示;
所述醇脱氢酶的氨基酸序列如SEQ ID NO:9所示;
所述甲酸脱氢酶的氨基酸序列如SEQ ID NO:10所示。
进一步地,编码6-磷酸葡萄糖酸脱氢酶的基因的核苷酸序列如SEQ ID NO:2所示;
编码葡萄糖脱氢酶的基因的核苷酸序列如SEQ ID NO:3所示;
编码醇脱氢酶的基因的核苷酸序列如SEQ ID NO:4所示;
编码甲酸脱氢酶的基因的核苷酸序列如SEQ ID NO:5所示。
一种重组细胞,所述重组细胞内所述的转化四氢姜黄素用核酸构建体。
一种转化四氢姜黄素用核酸构建体的应用,所述的一种转化四氢姜黄素用核酸构建体在姜黄素转化为四氢姜黄素的生产中的应用。
一种四氢姜黄素的制备方法,包括以下步骤:
S1,将所述的转化四氢姜黄素用核酸构建体载入到宿主细胞中得到重组细胞,然后对所述重组细胞培养并诱导表达,得到催化酶液;
S2,将所述催化酶液与姜黄素和脱氢酶的反应底物混合并进行全细胞催化,生成四氢姜黄素。
实施例1
姜黄素还原酶和用于还原辅酶的脱氢酶的制备
1)姜黄素还原酶yncB的克隆
使用以下引物,从大肠杆菌BL21(DE3)中克隆出姜黄素还原酶yncB:
上游引物:5′-CGCGGATCCATGGGGCAACAAAAGCAGCG-3′,
下游引物:5′-CCGCTCGAGTTAATCATCACCCGCCACGCGG-3′。
然后借助克隆试剂盒pMDTM18-T Vector Cloning Kit将所得的姜黄素还原酶yncB片段插入到载体pMD-18-T中。
在成功转化该构建体后,使用以下引物克隆出yncB:
上游引物:
5′-CAGCAAATGGGTCGCGGATCCATGGGGCAACAAAAG-3′
下游引物:5′-GTGGTGGTGGTGGTGCTCGAGTTAATCATCACCCGC-3′
借助Uniclone One Step Seamless Cloning Kit(一步法无缝克隆试剂盒)将所得yncB同源重组到pET-28a载体中,构建pET-28a-yncB质粒;然后将所述质粒引入大肠杆菌BL21(DE3)中并在此成功表达。成功转化后,借助测序确定序列同一性,其中编码所述yncB还原酶基因的核苷酸序列如SEQ ID NO:1所示,所述姜黄素还原酶的氨基酸序列如SEQ IDNO:6所示。
2)还原辅酶的脱氢酶的获取
本实施例中用于还原辅酶的脱氢酶中包括:葡萄糖脱氢酶(GDH)、6-磷酸葡萄糖酸脱氢酶(6PGDH)、甲酸脱氢酶(FDH)、醇脱氢酶(ADH),上述所述还原辅酶的脱氢酶由通用生物(安徽)股份有限公司基因合成获得。其中,编码6-磷酸葡萄糖酸脱氢酶的基因的核苷酸序列如SEQ ID NO:2所示;
编码葡萄糖脱氢酶的基因的核苷酸序列如SEQ ID NO:3所示;
编码醇脱氢酶的基因的核苷酸序列如SEQ ID NO:4所示;
编码甲酸脱氢酶的基因的核苷酸序列如SEQ ID NO:5所示。
所述6-磷酸葡萄糖酸脱氢酶的氨基酸序列如SEQ ID NO:7所示;
所述葡萄糖脱氢酶的氨基酸序列如SEQ ID NO:8所示;
所述醇脱氢酶的氨基酸序列如SEQ ID NO:9所示;
所述甲酸脱氢酶的氨基酸序列如SEQ ID NO:10所示。
实施例2
菌株BY4742/pESC-HIS-yncB-6PGDH的构建
包括以下步骤:
1)采用试剂盒HiPure Plasmid Micro Kit对实施例1中姜黄素还原酶yncB克隆体和6PGDH酶进行提取,得到pET-28a-yncB质粒、pET-32a-6PGDH质粒。
2)将pET-28a-yncB质粒通过对比BamHⅠ和XhoⅠ进行酶切获得姜黄素还原酶yncB的基因片段;将pET-32a-6PGDH质粒通过EcoRⅠ和NotⅠ进行酶切获得6PGDH酶的基因片段;对pESC-HIS通过BamHⅠ、XhoⅠ、EcoRⅠ和NotⅠ进行酶切获得pESC-HIS载体。
3)借助T4 DNA连接酶将yncB片段和6PGDH片段连接到pESC-HIS载体上,形成pESC-HIS-yncB-6PGDH构建体,所述pESC-HIS-yncB-6PGDH构建体的质粒谱图如图1所示;然后将pESC-HIS-yncB-6PGDH构建体引入到酿酒酵母菌BY4742中并在此成功表达。成功转化后,借助测序确定序列同一性,得到菌株BY4742/pESC-HIS-yncB-6PGDH。
实施例3
菌株BL21/pET-28a-yncB-6PGDH的构建
包括以下步骤:
1)采用试剂盒HiPure Plasmid Micro Kit对实施例1中姜黄素还原酶yncB得到pET-28a-yncB质粒;然后通过XhoⅠ进行酶切,得到yncB的基因片段。
2)使用以下引物克隆出6-磷酸葡萄糖酸脱氢酶(6PGDH):
上游引物:5′-GTGGCGGGTGATGATTAATTTGTTTAACTTTAAGAA-3′,
下游引物:
5′-GTGGTGGTGGTGCTCGAGTGCATTTGCCGGTTCTCT-3′;
借助Uniclone One Step Seamless Cloning Kit将6PGDH片段同源重组到酶切后的pET-28a-yncB中,得到pET-28a-yncB-6PGDH,所述pET-28a-yncB-6PGDH构建体的质粒谱图如图2所示;然后将pET-28a-yncB-6PGDH构建体引入大肠杆菌BL21(DE3)中并在此成功表达。成功转化后,借助测序确定序列同一性,得到菌株BL21/pET-28a-yncB-6PGDH。
实施例4
一种四氢姜黄素的制备方法,包括以下步骤:
取实施例2的BY4742/pESC-HIS-yncB-6PGDH菌液以1%接种量接种到SD-△his种子培养基中,30℃200rpm培养1天,培养结束,离心收集菌体,菌体转移至SD-△his诱导培养基中,30℃200rpm培养5天。诱导结束,离心收集菌体,用pH 6.0磷酸钾缓冲液重悬浮成菌液,进行全细胞催化。全细胞催化结束,反应液离心,分离上清和沉淀,取沉淀加入等量DMSO重悬浮,再次离心,取上清进行HPLC检测。
同时对姜黄素的反应前后进行外观评测,如图3-4所示,反应后的物料的颜色从橙色变成淡黄色,证明本实施例的菌液对姜黄素具有良好的转化作用。
其中,姜黄素生物转化为四氢姜黄素的反应式如式Ⅰ所示:
其中,全细胞催化体系:1g/L姜黄素、200g/L菌液、2g/L葡萄糖,反应缓冲液为pH6.0磷酸钾缓冲液;
催化条件:30℃,200rpm,24h。
培养基配方:
100mL SD-△his种子培养基:84mL灭菌水+10mL 20%葡萄糖+5mL 1×YNB+1mL100×AA(△his)溶液。
200mL SD-△his诱导培养基:164mL灭菌水+24mL半乳糖+10mL 1×YNB+2mL 100×AA(△his)溶液。
100×AA(△his)溶液:0.25g亮氨酸+0.05g赖氨酸+0.05g尿嘧啶配制成25mL混合水溶液,过滤除菌。
166.67g/L半乳糖:称量6.25g半乳糖配制成37.5mL水溶液,过滤除菌。
1×YNB溶液:称量5.36g YNB和11.76g硫酸铵,配制成157.6mL水溶液,过滤除菌。
实施例5
一种四氢姜黄素的制备方法,包括以下步骤:
取实施例3的BL21/pET-28a-yncB-6PGDH菌液以1%接种量接种到种子培养基中,37℃200rpm过夜培养,作为种子液。种子液接种到2L发酵培养基中,37℃200rpm培养至菌体对数生长期,加入0.4mM IPTG,20℃诱导18h。诱导结束,低温离心收集菌体,用pH6.0 Tris-HCl重悬浮成菌液,进行全细胞催化。全细胞催化结束,反应液离心,分离上清和沉淀,取沉淀加入等量DMSO重悬浮,再次离心,取上清进行HPLC检测。
其中,全细胞催化体系:1g/L姜黄素、200g/L菌液、2g/L葡萄糖,反应缓冲液为pH6.0 Tris-HCl;
催化条件:30℃,200rpm,24h。
种子培养基:40g/L葡萄糖、2g/L硫酸铵、2g/L磷酸二氢铵、1g/L七水合硫酸镁、20g/L酵母提取物、2g/L玉米浆干粉、80mg/L七水合硫酸亚铁、80mg/L一水合硫酸锰,高温高压115℃灭菌20min,使用前加入0.1%硫酸卡那霉素,并通过流加25%氨水调节pH至7.0。
发酵培养基:40g/L葡萄糖、1.8g/L硫酸铵、3g/L磷酸二氢铵、2g/L七水合硫酸镁、1g/L酵母提取物、2g/L玉米浆干粉、80mg/L七水合硫酸亚铁、80mg/L一水合硫酸锰,高温高压115℃灭菌20min,使用前加入0.1%硫酸卡那霉素,并通过流加25%氨水调节pH至7.0。
实施例6
一种四氢姜黄素的制备方法,本实施例与实施例4不同点仅在于:将6-磷酸葡萄糖酸脱氢酶(6PGDH)替换为葡萄糖脱氢酶(GDH)。
本实施例的姜黄素生物转化为四氢姜黄素的反应式如式Ⅱ所示:
实施例7
一种四氢姜黄素的制备方法,本实施例与实施例4不同点仅在于:将6-磷酸葡萄糖酸脱氢酶(6PGDH)替换为甲酸脱氢酶(FDH);同时将所述全细胞催化体系中的葡萄糖替换为等量的甲酸钠。
本实施例的姜黄素生物转化为四氢姜黄素的反应式如式Ⅲ所示:
实施例8
一种四氢姜黄素的制备方法,本实施例与实施例4不同点仅在于:将6-磷酸葡萄糖酸脱氢酶(6PGDH)替换为甲酸脱氢酶(FDH),同时将所述全细胞催化体系中的葡萄糖替换为等量的异丙醇。
本实施例的姜黄素生物转化为四氢姜黄素的反应式如式Ⅳ所示:
实施例9
取实施例4和实施例5的上清液进行姜黄素和四氢姜黄素的HPLC检测;
HPLC检测方法具体为:流动相A:乙酸氨:0.15%乙酸,用氨水调节pH到9.5;流动相B:100%乙腈;其他条件:色谱柱:Xtimate C18(4.6×250mm,5μm);检测波长:290nm,流速1mL/min;柱温为30℃;进样量10μL,等梯度洗脱:50%乙酸氨:50%乙腈。
全细胞催化产物检测结果见图5和图6,四氢姜黄素标准品HPLC检测结果见图7。
如图5和图6所示,全细胞催化后,该HPLC检测结果图中,在7.116 min和7.098 min有出峰,与四氢姜黄素标准品出峰时间一致,并且在样品中没有检出姜黄素,所以姜黄素已完全消耗,全部转化为四氢姜黄素;证明本发明的实施例2构建的BY4742/pESC-HIS-yncB-6PGDH和实施例3构建的BL21/pET-28a-yncB-6PGDH均能实现姜黄素生物转化为四氢姜黄素,转化率达到100%。
上述实施方式仅为本发明的优选实施方式,不能以此来限定本发明保护的范围,本领域的技术人员在本发明的基础上所做的任何非实质性的变化及替换均属于本发明所要求保护的范围。
Claims (10)
1.一种转化四氢姜黄素用核酸构建体,其特征在于,包括编码姜黄素还原酶的基因和编码用于还原辅酶的脱氢酶的基因。
2.根据权利要求1所述的一种转化四氢姜黄素用核酸构建体,其特征在于,所述姜黄素还原酶包括如下蛋白质(a)或(b):
(a)由氨基酸序列如SEQ ID NO:6所示的蛋白质;
(b)在(a)限定的氨基酸序列中经过取代、缺失或添加一种或几个氨基酸且具有姜黄素还原酶活性的由(a)衍生的具有至少95%序列同一性的同源蛋白质。
3.根据权利要求1或2所述的一种转化四氢姜黄素用核酸构建体,其特征在于:所述姜黄素还原酶包括curA还原酶、yncB还原酶、yfeF还原酶、ybjS还原酶和ygfF还原酶中至少一种。
4.根据权利要求3所述的一种转化四氢姜黄素用核酸构建体,其特征在于:编码所述yncB还原酶基因的核苷酸序列如SEQ ID NO:1所示。
5.根据权利要求1所述的一种转化四氢姜黄素用核酸构建体,其特征在于:所述还原辅酶的脱氢酶包括6-磷酸葡萄糖酸脱氢酶、葡萄糖脱氢酶、醇脱氢酶和甲酸脱氢酶中的至少一种。
6.根据权利要求5所述的一种转化四氢姜黄素用核酸构建体,其特征在于:
所述6-磷酸葡萄糖酸脱氢酶的氨基酸序列如SEQ ID NO:7所示;
所述葡萄糖脱氢酶的氨基酸序列如SEQ ID NO:8所示;
所述醇脱氢酶的氨基酸序列如SEQ ID NO:9所示;
所述甲酸脱氢酶的氨基酸序列如SEQ ID NO:10所示。
7.根据权利要求6所述的一种转化四氢姜黄素用核酸构建体,其特征在于:
编码6-磷酸葡萄糖酸脱氢酶的基因的核苷酸序列如SEQ ID NO:2所示;
编码葡萄糖脱氢酶的基因的核苷酸序列如SEQ ID NO:3所示;
编码醇脱氢酶的基因的核苷酸序列如SEQ ID NO:4所示;
编码甲酸脱氢酶的基因的核苷酸序列如SEQ ID NO:5所示。
8.一种重组细胞,其特征在于:所述重组细胞内含有如权利要求1-7任一项所述的转化四氢姜黄素用核酸构建体。
9.一种转化四氢姜黄素用核酸构建体的应用,其特征在于:权利要求1-7任一项所述的一种转化四氢姜黄素用核酸构建体在姜黄素转化为四氢姜黄素的生产中的应用。
10.一种四氢姜黄素的制备方法,其特征在于,包括以下步骤:
S1,将权利要求1-7任一项所述的转化四氢姜黄素用核酸构建体载入到宿主细胞中得到重组细胞,然后对所述重组细胞培养并诱导表达,得到催化酶液;
S2,将所述催化酶液与姜黄素和脱氢酶的反应底物混合并进行全细胞催化,生成四氢姜黄素。
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