Purification methods for recombinant Lactobacillus casei thymidylate synthase and mutants: a general, automated procedure.

Abstract:

:General procedures for the rapid, large-scale purification of recombinant Lactobacillus casei thymidylate synthase and its mutants have been established. An effective method employs sequential phosphocellulose and hydroxylapatite chromatography. Crude cell extracts are directly applied to phosphocellulose, and the enzyme is obtained in a nearly pure state by stepwise elution with KCl. The eluate is directly applied to hydroxylapatite, and the homogeneous enzyme is eluted with a gradient of potassium phosphate. The method has been successful for the purification of recombinant wild-type enzyme and all mutants thus far examined. The entire purification procedure has been automated using a commonly available FPLC system and can be performed in several hours with minimal operator time.

journal_name

Protein Expr Purif

authors

Kealey JT,Santi DV

doi

10.1016/s1046-5928(05)80039-7

keywords:

subject

Has Abstract

pub_date

1992-10-01 00:00:00

pages

380-5

issue

5

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(05)80039-7

journal_volume

3

pub_type

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