The isolation and characterization of phenylalanine ammonia lyase from Schizophyllum commune and an attempt to isolate the gene

Author

Jiamin Zeng

Keywords and Phrases

Schizophyllum commune

Abstract

"Phenylalanine ammonia-lyase (PAL) (EC 4. 3. 1. 5) activity was greatly increased in Schizophyllum commune 4-40 by growing the organism in a medium containing L-Phenylalanine. S. commune 4-40 PAL was purified 19.2 fold by a procedure utilizing Sephadex G-50 chromatography, ammonium sulfate fractionation and chromatographic separation on Sephadex G-200. The enzyme has a native molecular weight of 247,000. The K_m value for L-phenylalanine is 0.232 mM, the V_max is 0.415 mM/minute and the k_cat is 0.0164/second for the purified lyase.

Two attempts were made to synthesize a cDNA library from mRNA isolated from S. commune 4-40. Soybean PAL DNA and Rhodotorula rubra PAL DNAs were used as probes for screening for the PAL gene in the cDNA libraries and a genomic library prepared by Specht and Ulrich. The inability to cut the DNA isolated from reconstituted phage supposedly containing DNA from the cDNA library into more than 2 fragments with a mixture of two restriction endonucleases plus the high incidence of positive plaques in the screening tests, suggest that a true library had not been synthesized"--Abstract, page iii.

Advisor(s)

Siehr, Donald J.

Committee Member(s)

Hanna, Samir B.
Frank, Ronald L.

Department(s)

Chemistry

Degree Name

Ph. D. in Chemistry

Publisher

University of Missouri--Rolla

Publication Date

Summer 1993

Pagination

x, 111 pages

Note about bibliography

Includes bibliographical references (pages 107-110).

Rights

© 1993 Jiamin Zeng, All rights reserved.

Document Type

Dissertation - Restricted Access

File Type

text

Language

English

Subject Headings

PhenylalanineDNA -- AnalysisMessenger RNA

Thesis Number

T 6634

Print OCLC #

29302030

Electronic OCLC #

905245859

Recommended Citation

Zeng, Jiamin, "The isolation and characterization of phenylalanine ammonia lyase from Schizophyllum commune and an attempt to isolate the gene" (1993). Doctoral Dissertations. 1019.
https://scholarsmine.mst.edu/doctoral_dissertations/1019