Histidine

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: The study is well conducted and documented, however no OECD guideline was followed and the study is not GLP

Data source

Reference

Reference Type:

publication

Title:

Mutagenesis by normal metabolites in Escherichia coli: phenylalanine mutagenesis is dependent on error-prone DNA repair

Author:

Sargentini and Smith

Year:

1986

Bibliographic source:

Mutation Research, 161: 113-118

Materials and methods

Principles of method if other than guideline:

19 amino acids were tested for mutagenicity in E. coli K-12 uvrB DNA repair-deficient strain. The specific goal was finding a type of mutagenesis that is regulated by DNA repair genes.

GLP compliance:

no

Type of assay:

other: DNA repair assay in bacteria

Test material

Test material information

Constituent 1

Reference substance name:

Histidine

EC Number:

200-745-3

EC Name:

Histidine

Cas Number:

71-00-1

Molecular formula:

C6H9N3O2

IUPAC Name:

histidine

Test material form:

other: liquid

Details on test material:

Source: Pierce

Method

Target gene:

uvrB gene, which functions in the error-free excision repair of many bulky mutagenic lesions

Species / strain

Species / strain / cell type:

E. coli, other: K-12 strain

Details on mammalian cell type (if applicable):

DNA repair-deficient strains: SR250, SR 251, SR256 and SR1034.
Wild type strain: SR248

Test concentrations with justification for top dose:

2 mM

Controls

Untreated negative controls:

yes

Details on test system and experimental conditions:

19 amino acids were individually tested for their ability to revert lacZ53 cells to Lac+
Logarithmic-phase cells were prepared by diluting overnight cultures 1:50 and shaking the cultures at 37°C until a concentration of 4 x 10e8 colony-forming units (CFU) per mL was reached. The cells were pelleted and resuspende at 1.5 CFU/mL.
Cells were spread on Glu-600 plates (600 µg/mL glucose) or Glu-0 plates. On the GLu-600 plates 2 mM of the test compound was added.
Plates were incubated for 3 days at 37°C, after which the Lac+ mutants were scored.
The mean number of mutants was determined and the ration test/control was calculated.
Mean number of preexisting mutants per plate were: 1 (wild type), 15 (uvrB and 0 (uvrB umuC).
Mean number of plate mutants per control plate were: 60 (wild-type), 355 (uvrB), 21 (uvrB umuC).

Evaluation criteria:

2 criteria: The mean mutant frequency +/- 1 SD (range) for the amnio-acid-supplemented plates did not overlap the range for the control plates, and the mean relative (test/control) range for the uvrB strain did not overlap the mean relative range for the uvrB umuC strain.

Statistics:

The mutagenesis was normalized for constant growth.

Results and discussion

Test results

Species / strain:

E. coli, other: K-12 strain

Metabolic activation:

not applicable

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Untreated negative controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Any other information on results incl. tables

Effect of amino acids on spontaneous mutagenesis in E. coli

Values are the means of relative mutants per total cells data (mutant frequency on test plates divided by mutant frequency on control plates) from triplicate plate-assay experiments for LacZ53 strains.

Amino acid tested	Relative Lac^+mutagenesis in the presence of 2 mM
	uvrB	uvrB umuC
Histidine	0.93	1.04

Values did not indicate a significant effect on mutagenesis by either of two criteria.

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative

L-histidine did not show a significant effect on Lac reversion in the uvrB strain, and thus, was found not to bu mutagenic.

Executive summary:

In this study, 19 amino acids were tested to see if their catabolism was mutagenic, using uvrB cells. More specifically, the goal was to find a type of mutagenesis that is regulated by DNA repair genes. The study was not GLP and no OECD guideline was followed. A plate assay for mutagenesis was performed in which E. coli strains were incubated with the test substance for 3 days at 37°C with or without glucose present. Hereafter, Lac+ mutants were scored and the ratio test/control was determined, taking into account the growth-normalization factors.

L-histidine did not show a significant effect on Lac reversion in the uvrB strain, and thus, was found not to be mutagenic.