<?xml version="1.0" encoding="UTF-8"?><xml><records><record><source-app name="Biblio" version="7.x">Drupal-Biblio</source-app><ref-type>17</ref-type><contributors><authors><author><style face="normal" font="default" size="100%">Navale, Govinda R</style></author><author><style face="normal" font="default" size="100%">Sharma, Poojadevi</style></author><author><style face="normal" font="default" size="100%">Said, Madhukar S</style></author><author><style face="normal" font="default" size="100%">Ramkumar, Sudha</style></author><author><style face="normal" font="default" size="100%">Dharne, Mahesh S</style></author><author><style face="normal" font="default" size="100%">Thulasiram, H., V</style></author><author><style face="normal" font="default" size="100%">Shinde, Sandip S</style></author></authors></contributors><titles><title><style face="normal" font="default" size="100%">Enhancing epi-cedrol production in escherichia coli by fusion expression of farnesyl pyrophosphate synthase and epi-cedrol synthase</style></title><secondary-title><style face="normal" font="default" size="100%">Engineering in Life Science </style></secondary-title></titles><dates><year><style  face="normal" font="default" size="100%">2019</style></year><pub-dates><date><style  face="normal" font="default" size="100%">JULY</style></date></pub-dates></dates><language><style face="normal" font="default" size="100%">eng</style></language><abstract><style face="normal" font="default" size="100%">&lt;p&gt;Terpene &lt;span class=&quot;hitHilite&quot;&gt;synthase&lt;/span&gt; catalyses acyclic diphosphate &lt;span class=&quot;hitHilite&quot;&gt;farnesyl&lt;/span&gt; diphosphate into desired sesquiterpenes. &lt;span class=&quot;hitHilite&quot;&gt;In&lt;/span&gt; this study, a &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; enzyme was constructed &lt;span class=&quot;hitHilite&quot;&gt;by&lt;/span&gt; linking Santalum album &lt;span class=&quot;hitHilite&quot;&gt;farnesyl&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;pyrophosphate&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;synthase&lt;/span&gt; (SaFPPS) individually &lt;span class=&quot;hitHilite&quot;&gt;with&lt;/span&gt; terpene &lt;span class=&quot;hitHilite&quot;&gt;synthase&lt;/span&gt; and Artemisia annua &lt;span class=&quot;hitHilite&quot;&gt;Epi-cedrol&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;synthase&lt;/span&gt; (AaECS). &lt;span class=&quot;hitHilite&quot;&gt;The&lt;/span&gt; stop codon at &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; N-terminus &lt;span class=&quot;hitHilite&quot;&gt;of&lt;/span&gt; SaFPPS was removed and replaced &lt;span class=&quot;hitHilite&quot;&gt;by&lt;/span&gt; a short peptide (GSGGS) to introduce a linker between &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; two open reading frames. This &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; clone was expressed &lt;span class=&quot;hitHilite&quot;&gt;in&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;Escherichia&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;coli&lt;/span&gt; Rosseta DE3 cells. &lt;span class=&quot;hitHilite&quot;&gt;The&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; enzyme FPPS-ECS produced sesquiterpene 8-&lt;span class=&quot;hitHilite&quot;&gt;epi-cedrol&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;from&lt;/span&gt; substrates isopentenyl &lt;span class=&quot;hitHilite&quot;&gt;pyrophosphate&lt;/span&gt; and dimethylallyl &lt;span class=&quot;hitHilite&quot;&gt;pyrophosphate&lt;/span&gt; through sequential reactions. &lt;span class=&quot;hitHilite&quot;&gt;The&lt;/span&gt; K-m values &lt;span class=&quot;hitHilite&quot;&gt;for&lt;/span&gt; FPPS-ECS &lt;span class=&quot;hitHilite&quot;&gt;for&lt;/span&gt; isopentyl diphosphate was 4.71 mu M. &lt;span class=&quot;hitHilite&quot;&gt;The&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; enzyme carried out &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; efficient conversion &lt;span class=&quot;hitHilite&quot;&gt;of&lt;/span&gt; IPP to &lt;span class=&quot;hitHilite&quot;&gt;epi-cedrol&lt;/span&gt;, &lt;span class=&quot;hitHilite&quot;&gt;in&lt;/span&gt; comparison to single enzymes SaFPPS and AaECS when combined together &lt;span class=&quot;hitHilite&quot;&gt;in&lt;/span&gt; enzyme assay over time. Further, &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; recombinant E. &lt;span class=&quot;hitHilite&quot;&gt;coli&lt;/span&gt; BL21 strain harbouring &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; plasmid successfully produced &lt;span class=&quot;hitHilite&quot;&gt;epi-cedrol&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;in&lt;/span&gt; fermentation medium. &lt;span class=&quot;hitHilite&quot;&gt;The&lt;/span&gt; strain having &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; plasmid (pET32a-FPPS-ECS) produced 1.084 +/- 0.09 mg/L &lt;span class=&quot;hitHilite&quot;&gt;epi-cedrol&lt;/span&gt;, while &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; strain harbouring mixed plasmid (pRSETB-FPPS and pET28a-ECS) showed 1.002 +/- 0.07 mg/L titre &lt;span class=&quot;hitHilite&quot;&gt;in&lt;/span&gt; fermentation medium &lt;span class=&quot;hitHilite&quot;&gt;by&lt;/span&gt; overexpression and MEP pathway utilization. Structural analysis was done &lt;span class=&quot;hitHilite&quot;&gt;by&lt;/span&gt; I-TASSER server and docking was done &lt;span class=&quot;hitHilite&quot;&gt;by&lt;/span&gt; AutoDock Vina software, which suggested that secondary structure &lt;span class=&quot;hitHilite&quot;&gt;of&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; N- C terminal domain and their relative positions to functional domains &lt;span class=&quot;hitHilite&quot;&gt;of&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; enzyme was greatly significant to &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; catalytic properties &lt;span class=&quot;hitHilite&quot;&gt;of&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;the&lt;/span&gt; &lt;span class=&quot;hitHilite&quot;&gt;fusion&lt;/span&gt; enzymatic complex than individual enzymes.&lt;/p&gt;
</style></abstract><work-type><style face="normal" font="default" size="100%">Article</style></work-type><custom3><style face="normal" font="default" size="100%">&lt;p&gt;Foreign&lt;/p&gt;
</style></custom3><custom4><style face="normal" font="default" size="100%">&lt;p&gt;&lt;span class=&quot;info_value&quot;&gt;1.936&lt;/span&gt;&lt;/p&gt;
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