Thiaminase I of Bacillus thiaminolyticus

Douthit, H. A.; Airth, R. L.

doi:10.1016/0003-9861(66)90194-9

Cited by 26 publications

(20 citation statements)

References 14 publications

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“…Several alternative methods were developed [31], [32] before Edwin and Jackman [4] laid the foundation for a radiometric thiaminase assay that was used extensively for the next 40 years (see [18], [33], [34] for subsequent modifications of this method). The radiometric assay uses 14 C-thiamine as a substrate for thiaminase I, which releases thiazole -2- 14 C that is then extracted and the associated radioactivity is measured.…”

Section: Introductionmentioning

confidence: 99%

A Rapid Method for Assaying Thiaminase I Activity in Diverse Biological Samples

2014

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Vitamin B1 (thiamine) deficiencies can lead to neurological disorders, reproductive failure and death in wild and domestic animal populations. In some cases, disease is brought about by the consumption of foods high in thiaminase I activity. Levels of thiaminase activity in these foods are highly variable and the factors leading to production of this enzyme are poorly understood. Here we describe improvements in a spectrophotometric thiaminase I activity assay that measures the disappearance of 4-nitrothiophenol, a favored nucleophile co-substrate that replaces the thiazole portion of thiamine during the inactivation of thiamine by the enzyme. Scalable sample processing protocols and a 96-well microtiter plate format are presented that allow the rapid evaluation of multiple, replicated samples in the course of only a few hours. Observed levels of activity in bacterial culture supernatant, fish, ferns and molluscs using this colorimetric assay were similar to previously published reports that employed a radiometric method. Organisms devoid of thiaminase I, based upon previous work, showed no activity with this assay. In addition, activity was found in a variety of fishes and one fern species from which this enzyme had not previously been reported. Overall, we demonstrate the suitability of this technique for measuring thiaminase I activity within small amounts of tissue and environmental samples with replication levels that were heretofore prohibitive. The assay provides a considerable improvement in the ability to examine and understand the properties of an enzyme that has a substantial influence on organism and ecosystem health.

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Section: Introductionmentioning

confidence: 99%

A Rapid Method for Assaying Thiaminase I Activity in Diverse Biological Samples

2014

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“…More specifically, egg thiamine concentrations of lake trout feeding on high proportions of alewife were about one-half those of lake trout feeding on high proportions of rainbow smelt Osmerus mordax (Fitzsimons and Brown 1998). At the same time, Honeyfield et al (2002) reported on the isolation of thiaminase Iproducing bacteria, including Paenibacillus thiaminolyticus (formerly Bacillus thiaminolyticus; Douthit and Airth 1966;Nakamura 1990;Shida et al 1997) and other closely related Paenibacillaceae from frozen Lake Michigan alewife viscera. Taken together, these observations suggested the need for development of a new, more discriminating and optimized method to determine total thiaminase activity (optimized for thiaminase I) in tissue extracts of Great Lakes forage fishes and food chain organisms.…”

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confidence: 99%

A Method for Measuring Total Thiaminase Activity in Fish Tissues

et al. 2005

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An accurate, quantitative, and rapid method for the measurement of thiaminase activity in fish samples is required to provide sufficient information to characterize the role of dietary thiaminase in the onset of thiamine deficiency in Great Lakes salmonines. A radiometric method that uses 14 C-thiamine was optimized for substrate and co-substrate (nicotinic acid) concentrations, incubation time, and sample dilution. Total thiaminase activity was successfully determined in extracts of selected Great Lakes fishes and invertebrates. Samples included whole-body and selected tissues of forage fishes. Positive control material prepared from frozen alewives Alosa pseudoharengus collected in Lake Michigan enhanced the development and application of the method. The method allowed improved discrimination of thiaminolytic activity among forage fish species and their tissues. The temperature dependence of the thiaminase activity observed in crude extracts of Lake Michigan alewives followed a Q 10 ϭ 2 relationship for the 1-37ЊC temperature range, which is consistent with the bacterial-derived thiaminase I protein.

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“…Thereplacement of thiamine and glutamate by the yeast extract (5 g/ I ) in the IIledium of DOUTHIT and AIRTI3 [9] increased the production of extracellular thiaminase I by B. thiurninolyticus DBM 1068 (Tab. 1).…”

Section: Resultsmentioning

confidence: 99%