Functional Characterization of Novel Sesquiterpene Synthases from Indian Sandalwood, Santalum album.
Functional Characterization of Novel Sesquiterpene Synthases from Indian Sandalwood, Santalum album.
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DOI：
10.1038/srep10095
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发表时间：
2015-05-15
影响因子：
4.6
通讯作者：
Thulasiram HV
中科院分区：
文献类型：
--
作者：
Srivastava PL;Daramwar PP;Krithika R;Pandreka A;Shankar SS;Thulasiram HV
印度檀香因其芳香的心材油而备受重视，并以倍半萜的混合物为主。倍半萜烯是由金属依赖性萜烯环化酶催化法呢基二磷酸（FPP）环化而形成的。本研究利用转录组测序技术克隆了Sesquisabinene合成酶（SaSQS 1、SaSQS 2）、红没药烯合成酶（SaBS）、檀香烯合成酶（SaSS）和法呢基二磷酸合成酶（SaFDS）的5个基因，并对其功能进行了初步研究。专辑使用Illumina下一代测序，产生了3332万个高质量原始读数，其组装成84，094个单基因，平均长度为494.17 bp。在转录组测序的基础上，克隆了5个参与FPP、倍半卡宾烯、β-红没药烯和檀香烯生物合成的倍半萜烯降解酶SaFDS、SaSQS 1、SaSQS 2、SaBS和SaSS，并对其功能进行了初步研究。新的倍半萜烯脱氢酶（SaSQS 1和SaSQS 2）的特征在于倍半桧烯合酶的异构体具有不同的动力学参数和表达水平。此外，已经证明了在非优化的细菌细胞中从单基因SaSQS 1和SaSQS 2微生物生产倍半桧烯用于制备规模生产倍半桧烯的可行性。这些结果可能为檀香倍半萜在遗传上易处理的异源系统中的体内生产铺平了道路。
Indian Sandalwood, Santalum album L. is highly valued for its fragrant heartwood oil and is dominated by a blend of sesquiterpenes. Sesquiterpenes are formed through cyclization of farnesyl diphosphate (FPP), catalyzed by metal dependent terpene cyclases. This report describes the cloning and functional characterization of five genes, which encode two sesquisabinene synthases (SaSQS1, SaSQS2), bisabolene synthase (SaBS), santalene synthase (SaSS) and farnesyl diphosphate synthase (SaFDS) using the transcriptome sequencing of S. album. Using Illumina next generation sequencing, 33.32 million high quality raw reads were generated, which were assembled into 84,094 unigenes with an average length of 494.17 bp. Based on the transcriptome sequencing, five sesquiterpene synthases SaFDS, SaSQS1, SaSQS2, SaBS and SaSS involved in the biosynthesis of FPP, sesquisabinene, β-bisabolene and santalenes, respectively, were cloned and functionally characterized. Novel sesquiterpene synthases (SaSQS1 and SaSQS2) were characterized as isoforms of sesquisabinene synthase with varying kinetic parameters and expression levels. Furthermore, the feasibility of microbial production of sesquisabinene from both the unigenes, SaSQS1 and SaSQS2 in non-optimized bacterial cell for the preparative scale production of sesquisabinene has been demonstrated. These results may pave the way for in vivo production of sandalwood sesquiterpenes in genetically tractable heterologous systems.
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影响因子：
3.1
作者：
Aubourg, S;Lecharny, A;Bohlmann, J
通讯作者：
Bohlmann, J
DOI：
10.1111/j.1432-1033.1989.tb15000.x
发表时间：
1989-09-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
BEYER, P;MAYER, M;KLEINIG, H
通讯作者：
KLEINIG, H
影响因子：
56.9
作者：
KLAYMAN, DL
通讯作者：
KLAYMAN, DL
影响因子：
4.8
作者：
Jones, Christopher G.;Moniodis, Jessie;Bohlmann, Joerg
通讯作者：
Bohlmann, Joerg
影响因子：
4.3
作者：
Fujisawa, Masaki;Harada, Hisashi;Misawa, Norihiko
通讯作者：
Misawa, Norihiko