[14] δ-Aminolevulinic acid synthase (Rhodopseudomonas spheroides)

Burnham, Bruce F.

doi:10.1016/0076-6879(71)17179-0

Cited by 76 publications

(38 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Then the mixtures were heated in a water bath at 100°C for 15 min. After cooling for 15 min, 2 ml of the reaction mixture and 2 ml freshly prepared modified Ehrlich's reagent [22] were mixed together. The absorbance at 554 nm was measured at room temperature 30 min later.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Expression of a hemA Gene from Agrobacterium radiobacter in a Rare Codon Optimizing Escherichia coli for Improving 5-aminolevulinate Production

Lin

Chen

2008

Appl Biochem Biotechnol

View full text Add to dashboard Cite

The 5-aminolevulinate (ALA) synthase gene (hemA) from Agrobacterium radiobacter zju-0121, which was cloned previously in our laboratory, contains several rare codons. To enhance the expression of this gene, Escherichia coli Rosetta(DE3), which is a rare codon optimizer strain, was picked out as the host to construct an efficient recombinant strain. Cell extracts of the recombinant E. coli were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under the appropriate conditions. The results indicated that the activity of ALA synthase expressed in Rosetta(DE3)/pET-28a(+)-hemA was about 20% higher than that in E. coli BL21(DE3). Then the effects of precursors (glycine and succinate) and glucose, which is an inhibitor for ALA dehydratase as well as the carbon sources for cell growth, on the production of 5-aminolevulinate were investigated. Based on an optimal fed-batch culture system described in our previous work, up to 6.5 g/l (50 mM) ALA was produced in a 15-l fermenter.

show abstract

Section: Discussionmentioning

confidence: 99%

“…ALA was determined using modified Ehrlich's reagent [22]. Specifically, 2 ml of sample or standard was mixed with 1 ml 1.0 M sodium acetate (pH 4.6) in a cuvette, and 0.5 ml acetylacetone (2,4-pentanedione) was added to each cuvette.…”

Section: Discussionmentioning

confidence: 99%

Expression of a hemA Gene from Agrobacterium radiobacter in a Rare Codon Optimizing Escherichia coli for Improving 5-aminolevulinate Production

Lin

Chen

2008

Appl Biochem Biotechnol

View full text Add to dashboard Cite

show abstract

“…Then the mixtures were heated at 100 °C for 15 min. After cooling for 15 min, 2 ml of reaction mixture and 2 ml freshly prepared modified Ehrlich's reagent (Burnham, 1970) were mixed together. After 30 min, the absorbance at 554 nm was measured.…”

Section: Assay For Enzyme Activity and Alamentioning

confidence: 99%

High-level soluble expression of the hemA gene from Rhodobacter capsulatus and comparative study of its enzymatic properties

Lou

Zhu

et al. 2014

J. Zhejiang Univ. Sci. B

View full text Add to dashboard Cite

Abstract:The Rhodobacter capsulatus hemA gene, which encodes 5-aminolevulinic acid synthase (ALAS), was expressed in Escherichia coli Rosetta (DE3) and the enzymatic properties of the purified recombinant ALAS (RC-ALAS) were studied. Compared with ALASs encoded by hemA genes from Agrobacterium radiobacter (AR-ALAS) and Rhodobacter sphaeroides (RS-ALAS), the specific activity of RC-ALAS reached 198.2 U/mg, which was about 31.2% and 69.5% higher than those of AR-ALAS (151.1 U/mg) and RS-ALAS (116.9 U/mg), respectively. The optimum pH values and temperatures of the three above mentioned enzymes were all pH 7.5 and 37 °C, respectively. Moreover, RC-ALAS was more sensitive to pH, while the other two were sensitive to temperature. The effects of metals, ethylene diamine tetraacetic acid (EDTA), and sodium dodecyl sulfate (SDS) on the three ALASs were also investigated. The results indicate that they had the same effects on the activities of the three ALASs. SDS and metal ions such as Co

show abstract

“…These leaves differ conspicuously from untreated leaves in having absorption peaks at 552 and 590 nm. In addition, the absorption at 633 nm is higher than 9 that at 650 nm, indicating a greater proportion of pchld., the species of pchld which is not directly phototransformable to 6 chld (13,14,16,22), as compared to phototransformable pchld., the pchld species associated with the pchld holochrome 3 (13,14,16,22). Fivemin after irradiation of either control (not shown) or treated leaves with a 1-msec light flash, the absorption 0 at 650 nm has diminished, and the absorption peak at 683 nm associated with chld has appeared.…”

mentioning

confidence: 99%