Calcium alginate bead immobilization of cells containing tyrosine ammonia lyase activity for use in the production of p-hydroxycinnamic acid.
An Escherichia coli catalyst with tyrosine ammonia lyase activity (TAL) has been stabilized for repeated use in batch conversions of high tyrosine solids to p-hydroxycinnamic acid (pHCA). The TAL biocatalyst was stabilized by controlling the reaction pH to 9.8 +/- 0.1 and immobilizing the cells within a calcium alginate matrix that was cross-linked with glutaraldehyde and polyethyleneimine (GA/PEI). We found a GA range where the bead-encapsulated TAL was not inactivated, and the resulting cross-linking provided the beads with the mechanical stability necessary for repeated use in consecutive batch reactions with catalyst recycle. The GA/PEI calcium alginate TAL catalyst was used in 41 1-L batch reactions where 50 g L(-1) tyrosine was converted to 39 +/- 4 g L(-1) pHCA in each batch. The practical usefulness and ease of this process was demonstrated by scaling up the TAL bead immobilization and using the immobilized TAL catalyst in four 125-L bioconversion reactions to produce over 12 kg of purified pHCA.
Trotman
Robert J
RJ
Central Research and Development Department, E. I. du Pont de Nemours and Co., Experimental Station, Wilmington, Delaware 19880-0328, USA.
Camp
Carl E
CE
Ben-Bassat
Arie
A
DiCosimo
Robert
R
Huang
Lixuan
L
Crum
Grace A
GA
Sariaslani
F Sima
FS
Haynie
Sharon L
SL
D000464:Alginates / Q000737:chemistry*; D000642:Ammonia-Lyases / Q000378:metabolism*; D018914:Cells, Immobilized / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D003373:Coumaric Acids / Q000378:metabolism*; D004795:Enzyme Stability; D020723:Glucuronic Acid / Q000737:chemistry; D005976:Glutaral / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008863:Microspheres; D015394:Molecular Structure; D010636:Phenols / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D013696:Temperature; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism
D000464:Alginates; D003373:Coumaric Acids; D006603:Hexuronic Acids; D010636:Phenols; D014443:Tyrosine; D020723:Glucuronic Acid; D011094:Polyethyleneimine; D000642:Ammonia-Lyases; C023208:L-tyrosine ammonia-lyase; C030626:4-vinylphenol; D005976:Glutaral