Cloning and characterization of a novel β‐carotene hydroxylase gene from Lycium barbarum and its expression in Escherichia coli. (16th May 2014)

• Record Type: Journal Article
• Authors: Wu, Jiang; Ji, Jing; Wang, Gang; Li, Zhaodi; Diao, Jinjin; Wu, Guangxia
• Is Part Of: Biotechnology and applied biochemistry. Volume 61:Number 6(2014)
• Page Start: 637
• Page End: 645
• Publication Date: 2014-05-16
• Subjects: Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.1224
• Languages: English
• ISSNs: 0885-4513

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <p> <italic>Lycium barbarum</italic> contains high levels of zeaxanthin, which is produced by the conversion of <inline-formula><alternatives><inline-graphic mimetype="image" xlink:href="ark:/27927/pgh2wpwm3nh" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink" /><mml:math display="inline" altimg="urn:x-wiley:08854513:bab1224:equation:bab1224-math-0002" overflow="scroll" xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mi>β</mml:mi></mml:math></alternatives></inline-formula>‐carotene into zeaxanthin. <inline-formula><alternatives><inline-graphic mimetype="image" xlink:href="ark:/27927/pgh2wpwm3jv" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink" /><mml:math display="inline" altimg="urn:x-wiley:08854513:bab1224:equation:bab1224-math-0003" overflow="scroll" xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mi>β</mml:mi></mml:math></alternatives></inline-formula>‐Carotene hydroxylase catalyzes this reaction. We cloned a cDNA (<italic>chyb</italic>) encoding <inline-formula><alternatives><inline-graphic mimetype="image" xlink:href="ark:/27927/pgh2wpwm3kd" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink" /><mml:math display="inline" altimg="urn:x-wiley:08854513:bab1224:equation:bab1224-math-0004" overflow="scroll" xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mi>β</mml:mi></mml:math></alternatives></inline-formula>‐carotene hydroxylase (Chyb) from the <italic>L. barbarum</italic> leaf. A 939‐bp full‐length cDNA sequence was determined with 3′‐rapid amplification of cDNA end assay encoding a deduced Chyb protein (34.8 kDa) with a theoretical isoelectric point of 8.36. A bioinformatics analysis showed that the <italic>L. barbarum</italic> Chyb was located in the chloroplast. Further, to investigate the catalytic activity of the <italic>L. barbarum</italic> Chyb, a complementation analysis was conducted in <italic>Escherichia coli</italic>. The results strongly demonstrated that Chyb can catalyze <inline-formula><alternatives><inline-graphic mimetype="image" xlink:href="ark:/27927/pgh2wpwkzzx" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink" /><mml:math display="inline" altimg="urn:x-wiley:08854513:bab1224:equation:bab1224-math-0005" overflow="scroll" xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mi>β</mml:mi></mml:math></alternatives></inline-formula>‐carotene to produce zeaxanthin. Thus, this study suggests that <italic>L. barbarum</italic><inline-formula><alternatives><inline-graphic mimetype="image" xlink:href="ark:/27927/pgh2wpwm00g" xlink:type="simple" xmlns:xlink="http://www.w3.org/1999/xlink" /><mml:math display="inline" altimg="urn:x-wiley:08854513:bab1224:equation:bab1224-math-0006" overflow="scroll" xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mi>β</mml:mi></mml:math></alternatives></inline-formula>‐carotene hydroxylase could be a means of zeaxanthin production by genetic manipulation in <italic>E. coli</italic>.</p> </abstract>