A Method for the Purification of Coenzyme a from Yeast

Beinert, Helmut; Korff, R.W. Von; Green, D.E.; Buyske, D. A.; Handschumacher, Robert E.; Higgins, Harvey; Strong, F. M.

doi:10.1016/s0021-9258(18)38474-6

Cited by 47 publications

(6 citation statements)

References 25 publications

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“…7) agreed well with values of 0-65-0-71 jug. pantothenate/Lipmann unit reported for the pantothenate content of CoA (Lipmann et al 1947;Beinert et at. 1953).…”

Section: Coenzyme a Synthesismentioning

confidence: 99%

The synthesis of coenzyme A by Lactobacillus arabinosus 17–5

Pierpoint

Hughes

1954

Biochemical Journal

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I954 cation of the enzyme with an over-all yield of 20 % was effected.3. A purified preparation of the enzyme attacked all three phosphoproteins studied, but had no appreciable action on glycerophosphate or casein phosphopeptone. The enzyme is hence a true phosphoprotein phosphatase, distinct from the acid phosphomonoesterase.4. Optimum enzyme activity was found at pH 6-0 and at a substrate concentration corresponding to about 10,umoles/ml. of casein phosphorus with 0 001 M thioglycollic acid as activator.5. From the activation and inhibition studies it is deduced that sulphydryl and amino groups are essential for the activity of the enzyme. However the enzyme requires no dialysable coenzyme for its activity.The authors wish to thank the University of Madras for the award of a research studentship to one of us (T. A.S.) and for kind permission to publish the results which form part of a thesis approved for the degree of Master of Science.

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“…7) agreed well with values of 0-65-0-71 jug. pantothenate/Lipmann unit reported for the pantothenate content of CoA (Lipmann et al 1947;Beinert et at. 1953).…”

Section: Coenzyme a Synthesismentioning

confidence: 99%

The synthesis of coenzyme A by Lactobacillus arabinosus 17–5

Pierpoint

Hughes

1954

Biochemical Journal

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“…In parallel, these two teams also showed that CoA could be produced and isolated from microbial sources such as Streptomyces fradiae or yeast. , These observations represented an important advance for the study and production of CoA because the starting materials could be readily obtained (since the microorganisms could be grown easily in short time frames). Buyske and co-workers improved their process dramatically when they discovered that CoA could be coprecipitated with glutathione in the presence of cuprous sulfate . Using this strategy, they isolated 133 mg of CoA from 6 kg of dried yeast with a purity of ca.…”

Section: Isolation Of Coenzyme a From Microorganismsmentioning

confidence: 99%

“…Buyske and co-workers improved their process dramatically when they discovered that CoA could be coprecipitated with glutathione in the presence of cuprous sulfate. 15 Using this strategy, they isolated 133 mg of CoA from 6 kg of dried yeast with a purity of ca. 26%.…”

Section: ■ Introductionmentioning

confidence: 99%

Isolation and Synthesis of One of the Most Central Cofactors in Metabolism: Coenzyme A

Mouterde

Stewart

2018

Org. Process Res. Dev.

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The isolation and synthesis of Coenzyme A (CoA) has been an important field since this cofactor was discovered in 1947. CoA plays a central role in human metabolism and is vital in several metabolic pathways including fatty acid transport and degradation as well as the biosynthesis of a wide variety of compounds including fatty acids. The high cost of commercially-available CoA ($2600 / g with >85% purity) has motivated several research groups to find alternatives for its production. The variety of strategies that have been investigated for CoA production can be divided in three categories: isolation from microorganisms, total chemical synthesis and chemoenzymatic synthesis. These approaches provide access to CoA with different efficiencies. For example, direct isolation yields of ~25 mg/kg from dried yeast have been obtained. A variety of microorganisms such as Pseudomonas alkalytica, Sarcina lutea and Brevibacterium ammoniagenes accumulate CoA in their cultures at levels ranging from 0.03 to 115 mg/mL. Total chemical synthesis yields have ranged between 25 and 54% and chemoenzymatic approaches have provided overall yields of ca. 73%. This review covers all published for producing CoA in order to compare their efficiencies, scalabilities and convenience.

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“…Preparative quantities of CoASH have been isolated from microbial sources such as bakers' yeast 2,3 or Brevibacterium ammoniagenes IFO 12071. 4,5 Dried cells of wild-type or mutant B. ammoniagenes were subsequently used in chemo-enzymatic routes since they contained all five of the necessary CoASH biosynthetic enzymes.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Preparative quantities of CoASH have been isolated from microbial sources such as bakers’ yeast , or Brevibacterium ammoniagenes IFO 12071. , Dried cells of wild-type or mutant B. ammoniagenes were subsequently used in chemo-enzymatic routes since they contained all five of the necessary CoASH biosynthetic enzymes. , More recently, enzyme-assisted strategies have been developed by several groups for CoASH and its analogues − based on fundamental biochemical studies by Drueckhammer, , Strauss and Begley, − and Jackowski. − Our route to CoASH was inspired by the work of Strauss and Begley and employs the three-enzyme cascade of the CoASH salvage pathway (Scheme ). All three steps are ATP-dependent, and the proteins from Escherichia coli have been cloned and overexpressed. ,, Initial phosphorylation of 1 by pantetheine kinase (PanK) yields 2 , whose adenylation by phosphopantetheine adenyltransferase (PPAT) provides dephospho-CoA 3 .…”

Section: Introductionmentioning

confidence: 99%

An Efficient Chemoenzymatic Synthesis of Coenzyme A and Its Disulfide

Mouterde

Stewart

2016

Org. Process Res. Dev.

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We have developed a chemoenzymatic route to coenzyme A (CoASH) and its disulfide that is amenable to gram-scale synthesis using standard laboratory equipment. By synthesizing the symmetrical disulfide of pantetheine (pantethine), we avoided the need to mask the reactive sulfhydryl and also prevented sulfur oxidation by-products. No chromatography is required in our synthetic route to pantethine, which facilitates scale-up. Furthermore, we discovered that all three enzymes of the CoASH salvage pathway (pantetheine kinase, phosphopantetheine adenyltransferase and dephosphocoenzyme A kinase) accept the disulfide of the natural substrates and functionalize both ends of the molecules. This yields CoA disulfide as the product of the enzymatic cascade, a much more stable form of the cofactor. Free CoASH can be prepared by in situ S-S reduction.

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A Method for the Purification of Coenzyme a from Yeast

Cited by 47 publications

References 25 publications

The synthesis of coenzyme A by Lactobacillus arabinosus 17–5

The synthesis of coenzyme A by Lactobacillus arabinosus 17–5

Isolation and Synthesis of One of the Most Central Cofactors in Metabolism: Coenzyme A

An Efficient Chemoenzymatic Synthesis of Coenzyme A and Its Disulfide

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