• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.133-137
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• 1991

In order to increase the expression of XMP aminase [EC 6.3.4.1], which catalizes the conversion of 5'-XMP to the DNA fragment containing gua A gene coding for XMP aminase from pLC 34-10 plasmid was subcloned into pBR 322, and 1.7 kb gua A gene fragment was recloned under the control of trp promoter of pDR 720, E. coli expression vector. XMP aminase activity had increased by about 17 times when compared with that of the strain earring pLC 34-10.

• The Korean Journal of Mycology
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• v.21 no.4
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• pp.285-292
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• 1993

For the enzymatic conversion of 5'-XMP to 5'-GMP, partially purified XMP aminase from Escherichia coli was coupled with the yeast, Saccharomycrs cerevisiae, capable of ATP regeneration through glycolytic pathway. In order to elevate the level of XMP aminase in E. coli, $guaB^{-}(IMP\;dehydrogenase-less)$ mutant were introduced, and the yeast used as ATP supplier was treated by some method to increase its membrane permeability. The optimum conditions for efficient conversion reaction by energy-coupled system were investigated. As the results, a CH 41, $guaB^-$ mutant of E. coli K-12, showed 2.75 fold increase in the level of XMP aminase, compared with its parent cell. And the lyophylized yeast was the most effective at the ATP supplier. The optimum temperature and pH of conversion reaction were $40{\circ]C$ and pH 7.4, and the highest conversion ratio was shown under the reaction condition of 100 mM glucose, 100 mM inorganic phosphate and 6 mM AMP. When 36 units/ml XMP aminase used under the above conditions, the amount of 60 mg/ml yeast was sufficient to be used. Under the optimum condition, 71% of 1.8 mM(65.6 mg/100 ml) 5'-XMP was converted to 5'-GMP within 8 hr.

• Microbiology and Biotechnology Letters
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• v.7 no.3
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• pp.127-133
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• 1979

By the treatment of several mutagens, a number of 5'-guanylic acid producing mutants from 5'-xan-thylic acid were obtained from Brevibacterium ammoniagenes ATCC 6871. The indispensensable genetic-characters of the mutants were adenine requirement, lack of GMP-reductase and mutation to adenosine resistance from adenosine sensitiveness. Main product from 5'-xanthylic acirl by strain BA-17-2 was 5'-guanulic acid, and was isolated in a crystalline form by the use of anion exchange resin, Duolite 102 D. The isolated crystalline was identified as 5'-guanylic acid by means of paper chromatography, ultrav-iolet absorption spectra, and infrared absorption spectrum.

• Applied Biological Chemistry
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• v.24 no.2
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• pp.105-111
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• 1981

Growth responses of Brevibadterium ammoniagenes BA 17-2, which had been obtained by the treatment of several mutagens in our previous report, were investigated to select the preliminary optimal concentrations of phosphate, $Mg^{++}$, $Mn^{++}$ and thiamine for the production of 5'-XMP aminase. In this experiment it was shown that the concentration of phosphate in the medium has an important effect on the growth of microorganism. Using the medium containing 0.2% of $MgSO_4{\cdot}7H_2O$, 3mg/l of $MnSO_4{\cdot}H_2O$and $1\;mg/l$ of thiamine-HCl, the maximum cell mass was obtained at the concentration of 0.4% of $KH_2PO_4$ and $K_2HPO_4$, respectively. Above the concentration of these phosphates, cell growth was inhibited as the phosphate concentration increased to 1%, but the inhibition was overcome by the addition of 1% of $MgSO_4{\cdot}7H_2O$ and 3mg/l of thiamine-HCl. The 5'-XMP aminase activity was also influenced by the concentration of phosphate, $Mg^{++}$, $Mn^{++}$, and thiamine. In addition, the optimal culture pH and temperature for the enzyme activity were found to be 6.8 and $32^{\circ}C$, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.24 no.2
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• pp.111-116
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• 1991

The effective p.eduction of 5'-GMP(5'-Guanylic acid) by enzymatic conversion of 5'-XMP(5'-Xanthyic acid) was investigated. The Iyophilized Brevibacterium ammoniagenes ATCC 19216 which were used as the XHP aminase source, was immobilized by entrapping in K-carrageenan, agar, polyacrylamide or Ca-alginate. $3\%$ K-carrageenan was selected as the most suitable matrix. In the production of 5'-GMP using the free cells of 3. ammoniagenes ATCC 19216, the optimum conditions were $42^{\circ}C$, PH 7.0, 100mg/ml glucose, 120mg/ml cell ,8mg/ml $MgSO_4\cdot7H_2O$, 5mg/ml POESA, 5mg/ml phytic acid. Under the conditions, $94.5\%$ of 5'-GMP was converted within 8 hours. In the production of 5'-GMP using the immobilized whole cells of B. ammoniagenes ATCC 19216, the optimum conditions were $37^{\circ}C$, pH 7.5, 50mg/ml glucose, 1mg/ml $KH_2PO_4$, 10mg/ml phytic acid, 60mg/ml cell, 8mg/ml $MgSO_4\;\cdot\;7H_2O$, 5mg/ml POESA. Under the conditions, $64.7\%$ of 5'-GMP was converted within 40 hours.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1978.04a
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• pp.97.5-98
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• 1978

By the treatment of various mutagens, a number of 5'-guanylic acidproducing from 5'-xanthylic acid were obtained from Brevibacterium ammoniagenes ATCC 6871. The indispensable genetic characters of the mutants were adenine requirement, lack of GMP-reductase and mutation to adenosine resistance from adenosine sensitiveness. Main product of these mutants from 5'-xanthylic acid was 5'-guanylic acid. The substance was isolated in a crystalline form the culture broth of BA 17-2, and identified as 5'-guanylic acid by means of paper chromatography, ultra violet, absorption spectra, and infra red spectrum.