Biosynthesis, biotechnological production and applications of 5-aminolevulinic acid

Sasaki, Kohsuke; Watanabe, Masanori; Tanaka, Tohru

doi:10.1007/s00253-001-0858-7

Cited by 182 publications

(51 citation statements)

References 16 publications

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“…It is a precursor of tetrapyrroles and a key intermediate in regulating tetrapyrrole biosynthesis (Sasaki et al, 2002). ALA has been used in a number of applications, i.e., as a new kind of green pesticide in agriculture and as a photodynamic medicine for photodynamic diagnosis and therapy of cancer (Lang et al, 2001;Braathen et al, 2007;Berkovitch et al, 2008;Pariser et al, 2008;Zaak et al, 2008).…”

Section: Introductionmentioning

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

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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

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Section: Introductionmentioning

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

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“…Also, ALA as photosensitizer for photodynamic therapy is of potential use for other cancers [1,2], oral errucous hyperplasia [3], and so on. And ALA has received wide attention as a selective and biodegradable herbicide or insecticide [4,5] in agriculture.…”

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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

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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.

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“…For ALA batch and fed-batch fermentation, a single colony was inoculated into 5 ml BHIS medium and incubated at 180 rpm for 22 h at 30°C. The 5-ml preculture was then inoculated into a 300-ml baffled shake flask containing 50 ml modified CGXII medium; CGXII medium contained 20 g/liter (NH 4 The initial optical density at 600 nm (OD 600 ) was controlled at about 0.5, and the glucose concentration was controlled at 40 g/liter; 0.25 mM IPTG (isopropyl-␤-D-thiogalactopyranoside) was added at 0 h to induce the expression of plasmid genes. Fermentation was performed at 30°C and 180 rpm, and the pH was maintained at approximately 6.5 with 4 M NaOH throughout the fermentation.…”

Section: Methodsmentioning

confidence: 99%

“…ALA can also be used as a selective biodegradable herbicide and insecticide or an adversity resistance and growth-accelerating agent in agriculture (4,5).…”

mentioning

confidence: 99%

A New Strategy for Production of 5-Aminolevulinic Acid in Recombinant Corynebacterium glutamicum with High Yield

Yang

Liu

Cheng

et al. 2016

Appl Environ Microbiol

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5-Aminolevulinic acid (ALA), a nonprotein amino acid involved in tetrapyrrole synthesis, has been widely applied in agriculture, medicine, and food production. Many engineered metabolic pathways have been constructed; however, the production yields are still low. In this study, several 5-aminolevulinic acid synthases (ALASs) from different sources were evaluated and compared with respect to their ALA production capacities in an engineered Corynebacterium glutamicum CgS1 strain that can accumulate succinyl-coenzyme A (CoA). A codon-optimized ALAS from Rhodobacter capsulatus SB1003 displayed the best potential. Recombinant strain CgS1/pEC-SB produced 7.6 g/liter ALA using a mineral salt medium in a fed-batch fermentation mode. Employing two-stage fermentation, 12.46 g/liter ALA was produced within 17 h, with a productivity of 0.73 g/liter/h, in recombinant C. glutamicum. Through overexpression of the heterologous nonspecific ALA exporter RhtA from Escherichia coli, the titer was further increased to 14.7 g/liter. This indicated that strain CgS1/pEC-SB-rhtA holds attractive industrial application potential for the future. IMPORTANCEIn this study, a two-stage fermentation strategy was used for production of the value-added nonprotein amino acid 5-aminolevulinic acid from glucose and glycine in a generally recognized as safe (GRAS) host, Corynebacterium glutamicum. The ALA titer represented the highest in the literature, to our knowledge. This high production capacity, combined with the potential easy downstream processes, made the recombinant strain an attractive candidate for industrial use in the future.A s the common precursor of tetrapyrroles such as porphyrin, heme, vitamin B 12 , and chlorophyll, 5-aminolevulinic acid (ALA) has been reported to be effective in tumor-localizing and photodynamic therapy for various diseases (1-3). ALA can also be used as a selective biodegradable herbicide and insecticide or an adversity resistance and growth-accelerating agent in agriculture (4, 5).In living organisms, two kinds of metabolic pathways have been described for ALA biosynthesis (Fig. 1). One is the C 5 pathway, which occurs in algae, higher plants, and many bacteria, including Escherichia coli and archaea. The C 5 pathway involves the following three enzymatic activities: glutamyl-tRNA synthetase (GluRS) (encoded by gltX), a NADPH-dependent glutamyl-tRNA reductase (HemA, encoded by hemA), and a glutamate-1-semialdehyde aminotransferase (HemL, encoded by hemL). The other is the C 4 pathway, which is present in birds, mammals, yeast, and purple non-sulfur-photosynthetic bacteria. In this pathway, ALA is formed through one-step catalysis by 5-aminolevulinic acid synthase (ALAS), which condenses glycine and succinyl-coenzyme A (CoA), an intermediate of the tricarboxylic acid (TCA) cycle.In E. coli, the native pathway for ALA biosynthesis is the C 5 pathway, which is tightly regulated by feedback inhibition of the end product heme (6). Previously, we developed a strategy to produce ALA in recombinant E. coli vi...

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Biosynthesis, biotechnological production and applications of 5-aminolevulinic acid

Cited by 182 publications

References 16 publications

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

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

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

A New Strategy for Production of 5-Aminolevulinic Acid in Recombinant Corynebacterium glutamicum with High Yield

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