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

Phenylalanine
DNA -- Analysis
Messenger RNA

Thesis Number

T 6634

Print OCLC #

29302030

Electronic OCLC #

905245859

Link to Catalog Record

Electronic access to the full-text of this document is restricted to Missouri S&T users. Otherwise, request this publication directly from Missouri S&T Library or contact your local library.

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