Molecular cloning, expression, and biochemical characterization of carotenoid cleavage dioxygenase 1 (LbCCD1) from Lycium barbarum (Record no. 55540)

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fixed length control field 03572nam a2200313Ia 4500
003 - CONTROL NUMBER IDENTIFIER
control field MX-MdCICY
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20250626080925.0
040 ## - CATALOGING SOURCE
Transcribing agency CICY
090 ## - LOCALLY ASSIGNED LC-TYPE CALL NUMBER (OCLC); LOCAL CALL NUMBER (RLIN)
Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) B-12903
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245 10 - TITLE STATEMENT
Title Molecular cloning, expression, and biochemical characterization of carotenoid cleavage dioxygenase 1 (LbCCD1) from Lycium barbarum
490 0# - SERIES STATEMENT
Series statement Molecular Catalysis, 569, p.114561, 2024
500 ## - GENERAL NOTE
General note Artículo
520 3# - SUMMARY, ETC.
Summary, etc. Carotenoid cleavage dioxygenases (CCDs) play a pivotal role in the biosynthesis of volatile apocarotenoids, which have significant industrial applications due to their aromatic and bioactive properties. This study focuses on the molecular cloning and biochemical characterization of the LbCCD1 from Lycium barbarum. The LbCCD1 gene was successfully cloned and heterologously expressed in Escherichia coli and several carotenoids were using as substrates for investigate its specificity by in vitro and in vivo experiment. The LbCCD1 protein was able to cleave a variety of carotenoids including β-carotene, zeaxanthin, astaxanthin, and β-apo-8'-carotenal, at the 9, 10 (9', 10') double bond to produce β-ionone, 3‑hydroxy-4-oxo-β-ionone, and 3‑hydroxy-β-ionone, respectively in vitro. LbCCD1 could also cleave zeaxanthin and β-carotene at the 9, 10 (9', 10') double bond to produce β-ionone, respectively, in E. coli accumulating carotenoids. Interestingly, LbCCD1 did not exhibit cleavage activity on lycopene either in vivo or in vitro unlike other CCD1 family enzymes. In the previous experiment, it was confirmed that LbCCD1 exhibits cleavage activity towards β-apo-8'-carotenal in vitro, so we used β-apo-8'-carotenal as the substrate for characterizing the enzymatic properties. The expression of LbCCD1 was optimized at such conditions (temperature 24 ◦C, IPTG 0.1 mM, induction time 24 h). The biochemical characterization of LbCCD1 revealed the optimal activities were at pH 9 and 55 ◦C. The addition of 10 percent ethanol could increase enzyme activity to above 15 percent. However, the concentration of Fe2+ has a minimal effect on enzyme activity. The Vmax for β-apo-8'-carotenal was 8.6 U/mg, while the Km was 0.27 mM. To preliminarily verify the potential of LbCCD1 as a biological component for β-ionone production. By introducing LbCCD1 into the β-carotene-high-producing chassis cell and optimizing the conditions (temperature 30 ◦C, IPTG 0.01 mM, Fe2+ concentration 0.05 mM), the β-ionone yield reached 21.45 mg/L. This study focused on one of the CCDs derived from woody plants, which have been relatively underexplored. It lays the groundwork for expanding the CCD enzyme library, identifying suitable CCDs, and investigating the structure-function relationship of CCDs. Furthermore, it sets the stage for engineering novel CCD genes and developing advanced applications of CCDs as biocatalysts and platforms for synthetic biology. These advancements will enable the efficient production of volatile aroma compounds from carotenoids.
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element CAROTENOID CLEAVAGE DIOXYGENASES
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element CCD1 FAMILY
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element LYCIUM BARBARUM
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element BIOCHEMICAL CHARACTERIZATION
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element B-IONONE
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Ke, K.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Guo, L.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Qi, Z.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Dou, D.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Ma, H.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Fang, X.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Zhao, L.
856 40 - ELECTRONIC LOCATION AND ACCESS
Uniform Resource Identifier <a href="https://drive.google.com/file/d/1OYB6yyKaf5-4Dkbe5FgFUECtjmxLchCp/view?usp=drive_link">https://drive.google.com/file/d/1OYB6yyKaf5-4Dkbe5FgFUECtjmxLchCp/view?usp=drive_link</a>
Public note Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
942 ## - ADDED ENTRY ELEMENTS (KOHA)
Source of classification or shelving scheme Clasificación local
Koha item type Documentos solicitados
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  Clasificación local     Ref1 CICY CICY Documento préstamo interbibliotecario 26.06.2025   B-12903 26.06.2025 26.06.2025 Documentos solicitados