1993 Fiscal Year Final Research Report Summary
Biochemical degradation of phenols by yeast
| Project/Area Number |
03454031
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
都市工学・衛生工学
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| Research Institution | Yamanashi Univ. |
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Principal Investigator |
KATAYAMA Keiko Yamanashi Univ. Faculty of Eng. Research associate, 工学部, 助手 (40111778)
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| Co-Investigator(Kenkyū-buntansha) |
HIRAYAMA Kimiaki Yamanashi Univ. Faculty of Eng. Associate professor, 工学部, 助教授 (40107214)
KAZAMA Futaba Yamanashi Univ. Faculty of Eng. Research associate, 工学部, 助手 (00115320)
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| Project Period (FY) |
1991 – 1993
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| Keywords | Biodegration / Immobilization / Monochlorophenols / Phenol / Rhodotorula / Yeast |
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| Research Abstract |
Biodegradation of phenol and monochlorophenols (CPs) by a yeast strain of Rhodotorula glutinis was exmined. The strain completely degraded 5 mM of phenol and utilized phenol as a sole carbon source. The strain may degrade phenol by the "ortho" type of ring fission because muconolactone was observed in the cultured broth. 3-Chlorophenol (3-CP) and 4-chlorophenol (4-CP) were well degraded and stoichiometric release of chloride ion was observed, but degradation of 2-Chlorophenol was low with few amount of chloride ion. Biodegradability of 3-CP and 4-CP was increased by the addition of phenol or CPs to the medium at the cell cultivation. Since 4-chlorophenol and maleylacetic acid were determined as metabolites of 3-CP and 4-CP by GC/Ms analysis, dechlorination may take place between the formation of 4-chlorophenol and maleylacetic acid.
Biochemical degradation of phenols by immobilized yeast cells was examined. Repeated batch cultures (10 days) for the degradation of phenol were performed f
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or comparing three kinds of beads, made from cellulose, chitosan and sintered glass. In the initial incubation period, cellulose beads as a carrier had higher yeast cells immobilizing capacity and had higher removal of TOC and phenol(%) than chitosan and sintered glass beads. The numbers of immobilized yeast cells with cellulose beads finally reached 2.5 X 10^8 cells/ml-beads. This result was almost the same as with chitosan and sintered glass beads. Continuous culture was performed with cellulose beads. Phenol (5 mM) and 3- and 4-CP (0.05 mM) was degraded stably for 100 days in this system.
Identification of beta-ketoadipic acid in muconolactone and protocatechuic acid metabolites by the crude cell extract of yeast Rhodotorula rubra was carried out using methoxime-trimethylsilyl (MO-TMS) derivative to prevent enol ether formation. beta-Ketoadipic acid was converted successfully to MO-TMS derivative, and enol ether formation was not observed. Mass spectra of two isomers of MO-TMS derivative of beta-ketoadipic acid resembled each other in their fragmentation pattern ; they had sufficient intensities of molecular ion at m/z 333, and other characteristic ions. Less
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