Development of an improved in vitro activity assay for medium chain length PHA polymerases based on CoenzymeA release measurements

2000 ◽  
Vol 41 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Guy de Roo ◽  
Qun Ren ◽  
Bernard Witholt ◽  
Birgit Kessler
Keyword(s):  
Chain Length ◽  
Vitro Activity ◽  
In Vitro Activity ◽  
Activity Assay ◽  
Medium Chain ◽  
Medium Chain Length

scholarly journals Characterization of Site-Specific Mutations in a Short-Chain-Length/Medium-Chain-Length Polyhydroxyalkanoate Synthase:In VivoandIn VitroStudies of Enzymatic Activity and Substrate Specificity

2013 ◽  
Vol 79 (12) ◽  
pp. 3813-3821 ◽  
Author(s):  
Jo-Ann Chuah ◽  
Satoshi Tomizawa ◽  
Miwa Yamada ◽  
Takeharu Tsuge ◽  
Yoshiharu Doi ◽  
...  
Keyword(s):  
Chain Length ◽  
Fold Increase ◽  
Short Chain ◽  
Pha Synthase ◽  
Wild Type ◽  
Short Chain Length ◽  
Medium Chain ◽  
Medium Chain Length

ABSTRACTSaturation point mutagenesis was carried out at position 479 in the polyhydroxyalkanoate (PHA) synthase fromChromobacteriumsp. strain USM2 (PhaCCs) with specificities for short-chain-length (SCL) [(R)-3-hydroxybutyrate (3HB) and (R)-3-hydroxyvalerate (3HV)] and medium-chain-length (MCL) [(R)-3-hydroxyhexanoate (3HHx)] monomers in an effort to enhance the specificity of the enzyme for 3HHx. A maximum 4-fold increase in 3HHx incorporation and a 1.6-fold increase in PHA biosynthesis, more than the wild-type synthase, was achieved using selected mutant synthases. These increases were subsequently correlated with improved synthase activity and increased preference of PhaCCsfor 3HHx monomers. We found that substitutions with uncharged residues were beneficial, as they resulted in enhanced PHA production and/or 3HHx incorporation. Further analysis led to postulations that the size and geometry of the substrate-binding pocket are determinants of PHA accumulation, 3HHx fraction, and chain length specificity.In vitroactivities for polymerization of 3HV and 3HHx monomers were consistent within vivosubstrate specificities. Ultimately, the preference shown by wild-type and mutant synthases for either SCL (C4and C5) or MCL (C6) substrates substantiates the fundamental classification of PHA synthases.

Unmodified medium chain length polyhydroxyalkanoate (uMCL-PHA) as a thin film for tissue engineering application – characterization and in vitro biocompatibility

2015 ◽  
Vol 141 ◽  
pp. 55-58 ◽  
Author(s):  
Sangeetha Vasudevaraj Naveen ◽  
Irene Kit Ping Tan ◽  
Yuh Shan Goh ◽  
Hanumantha Rao Balaji Raghavendran ◽  
Malliga Raman Murali ◽  
...  
Keyword(s):  
Thin Film ◽  
Tissue Engineering ◽  
Chain Length ◽  
Engineering Application ◽  
Tissue Engineering Application ◽  
Medium Chain ◽  
In Vitro Biocompatibility ◽  
Medium Chain Length

Aliphatic α,γ-bis(Amides) of Methotrexate. Influence of Chain Length on In-vitro Activity Against Sensitive and Resistant Tumour Cells

1999 ◽  
Vol 5 (4) ◽  
pp. 299-305 ◽  
Author(s):  
R. Pignatello ◽  
G. Spampinato ◽  
V. Sorrenti ◽  
L. Vicari ◽  
C. Di Giacomo ◽  
...  
Keyword(s):  
Chain Length ◽  
Tumour Cells ◽  
Vitro Activity ◽  
In Vitro Activity

In Vitro Activity of Phytocannabinoids from High Potency Cannabis sativa

Planta Medica ◽  
2012 ◽  
Vol 78 (05) ◽  
Author(s):  
A Husni ◽  
S Ross ◽  
O Dale ◽  
C Gemelli ◽  
G Ma ◽  
...  
Keyword(s):  
Cannabis Sativa ◽  
Vitro Activity ◽  
In Vitro Activity ◽  
High Potency
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