Extraction and Activity of O-acetylserine(thiol)lyase (OASTL) from Microalga Chlorella sorokiniana.

Extraction and Activity of O-acetylserine(thiol)lyase (OASTL) from Microalga Chlorella sorokiniana.

Bio-protocol (2017-06-20)

Giovanna Salbitani, Simona Carfagna

ABSTRACT

O-acetylserine(thiol)lyase (OASTL) is an enzyme catalysing the reaction of inorganic sulphide with O-acetylserine to form the S-containing amino acid L-cysteine. Here we describe an improved protocol to evaluate the activity of this enzyme from the microalga Chlorella sorokiniana. It is a colorimetric assay based on the reaction between cysteine, the product of OASTL activity, and ninhydrin reagent, which forms a thiazolidine (Thz).

MATERIALS

Product Number

Brand

Product Description

451657

Sigma-Aldrich

Copper(II) sulfate, anhydrous, powder, ≥99.99% trace metals basis

E6760

Sigma-Aldrich

Ethylenediaminetetraacetic acid ferric sodium salt, BioReagent, suitable for plant cell culture, crystalline

S5886

Sigma-Aldrich

Sodium chloride, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%

D9779

Sigma-Aldrich

DL-Dithiothreitol, Molecular Biology, ≥98% (HPLC), ≥99% (titration)

H4034

Sigma-Aldrich

HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture

Z0251

Sigma-Aldrich

Zinc sulfate heptahydrate, BioReagent, suitable for cell culture

CDS020792

Sigma-Aldrich

O-Acetyl-L-serine

C5670

Sigma-Aldrich

Calcium chloride, anhydrous, BioReagent, suitable for insect cell culture, suitable for plant cell culture, ≥96.0%

M2643

Sigma-Aldrich

Magnesium sulfate, BioReagent, suitable for cell culture, suitable for insect cell culture

B6768

Sigma-Aldrich

Boric acid, BioReagent, Molecular Biology, suitable for cell culture, suitable for plant cell culture, ≥99.5%

P5655

Sigma-Aldrich

Potassium phosphate monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%