Lignin peroxidase , production

Nakamura Y, Sawada T, Sungusi MG, Kobayashi F, Kuwahara M, Ito H (1997) Lignin peroxidase production by Phanerochaete chrysosporium. J Chem Eng Jpn 30 1-6... 

While the batch process is the dominant one in current use, researchers and companies have attempted to create continuous bioreactor systems. Lopez et al. immobilized Candida rugosa in polymethacrylamide hydrazide beads and polyurethane foam 3 with the intent to achieve the continuous production of lipase enzymes. Despite flow problems with the polyurethane foam, it showed high lipolytic activity. Biomass buildup was problematic. Feijoo et al. immobilized Phanerochaete chry-sosporium on polyurethane foam in packed bed bioreactors under near-plug flow conditions. Continuous lignin peroxidase production was accomplished, the rate of which was studied as a function of recycle ratio. 

Cornwell RL, Tinland-Butez M-F, Tardone PJ, Cabasso I, Hammel KE. Lignin degradation and lignin peroxidase production in cultures of Phanerochaete chrysosporium immobilized on porous ceramic supports. Enzyme Microb Technol 1990 12 916-920. 

Ferrara, M. A., Bon, E. P. S., and Araujo Neto, J. S. 2002. Use of Steam Explosion Liquor from Sugar Cane Bagasse for Lignin Peroxidase Production by Phanero-chaete chrysosporium. Appl. Biochem. Biotechnol., 98-100,289-300. 

Zerbini, J. E., Oliveira, E. M. M., and Bon, E. P. S. 1999. Lignin peroxidase production by Streptomyces viridosporus T7A Nitrogen nutrition optimization using glucose as carbon source. Appl. Biochem. Biotechnol., 77-79, 681-688. 

Liebeskind M, Hoecker H, Wandrey C and Jaeger A G, Strategies for improved lignin peroxidase production in agitated pellet cultures of Phanerochaete chrysosporium and the use of a novel inducer , FEMS Microbiol. Lett., 1990, 71, 325-30. 

Perez J, TW Jeffries (1990) Mineralization of C-ring-labelled synthetic lignin correlates with the production of lignin peroxidase, not of manganese peroxidase or laccase. Appl Environ Microbiol 56 1806-1812. 

Van der Woude MW, K Boominathan, CA Reddy (1993) Nitrogen regulation of lignin peroxidase and manganese-dependent peroxidase production is independent of carbon and manganese regulation in Phanerochaete chrysosporium. Arch Microbiol 160 1-4. 

Pieper DH, R Winkler, H Sandermann (1992) Eormation of a toxic dimerization product of 3,4-dichloroani-line by lignin peroxidase from Phanerochaete chrysosporium. Angewandte Chemie 104 60-61. 

Bumpus JA, M Tatarko (1994) Biodegradation of 2,4,6-trinitrotoluene by Phanerochaete chrysosporium identification of initial degradation products and the discovery of a metabolite that inhibits lignin peroxidases. Curr Microbiol 28 185-190. 

Lentinula (Lentinus) edodes produced only Mn peroxidase, and the production of both laccase and lignin peroxidase was, apparently, negligible. Consider that a strict relation between the production of Mn peroxidase and the dye decolorization ability was observed in vivo [13]. 

Haapala A, Linko S (1993) Production of Phanerochaete chrysosporium lignin peroxidase under various culture conditions. Appl Microbiol Biotechnol 40 494-498... 

PUot-Scale Production and Properties of Lignin Peroxidases... 

Production of Lignin Peroxidase. Medium for the inoculum was rich in yeast extract (25 g/1) and glucose (25 g/l) to promote maximal growth of the mycelia. The inoculum of Phanerochaete chrysosporium ATCC 24725 was first cultivated for 3 days at 30°C in five litres of medium divided in five shake flasks. The shake flask batches were transferred to a 100 litre bioreactor and cultivated again for 3 days at 30°C. The batches were stirred and aerated to obtain maximal growth of mycelia. 

Figure 1. Pilot-scale production of lignin peroxidases...

Activity Assays. The standard activity assay mixture of 3 ml contained about 0.1 U/ml lignin peroxidase, 0.4 mM veratryl alcohol (Fluka, purum >97%) and 0.1M sodium tartrate, pH 3.0. The reaction was started by adding 15 fil of 54 mM H2O2 to make a final concentration of 0.28 mM in the reaction. The production of veratraldehyde was followed by recording the change of absorbance for 12 seconds at 310 nm in a cuvette which was thermostated to 37°C. The reaction was started 24 seconds before the recording. One unit of lignin peroxidase is defined as the amount of enzyme required to oxidize one imol of veratryl alcohol to veratraldehyde in one minute. 

Production. The inoculum grew vigorously in the rich yeast extract containing media and produced a thick viscous dispersion in the stirred tank bioreactor. No lignin peroxidase activity could be detected at this stage. When transferred to the 1000 1 production bioreactor, the mycelium of P.chrysosporium attached completely to the nylon wool sheets within a few hours after inoculation and the medium remained completely clear throughout the cultivation. The enzyme had to be harvested immediately after the maximal activity level was reached due to its... 

A detailed scientific study on the properties of the five major isozymic forms of the lignin peroxidases produced in our pilot reactor has recently been published 12), Our purified enzyme in this study is composed of two isozymes having isoelectric points of 3.85 and 3.80 and molecular masses of 42 000. In this study we have characterized the enzyme s stability as an industrial product. 

Figure 1. 3-0-4 lignin substructure model compounds and products of their degradation by ligninolytic cultures of Phanerochaete chrysosporium and by lignin peroxidase. D H, and 0 of 3-ether bond and of H2 0, respectively. B and C show results of stable isotope experiments. 

These findings led to the proposition that the veratryl alcohol is degraded via the quinone intermediates (Figure 5) to CO2 through a series of transformations involving lignin peroxidase, perhydroxy radicals and the NADP-dependent aryl alcohol oxidoreductase. Veratraldehyde, the major product of lignin peroxidase catalyzed veratryl alcohol oxidation, is rapidly reduced back to veratryl alcohol it is the further metabolism of the side products of the oxidative process, viz. the quinones and lactones, that drives the overall transformation towards completion (34). 

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