Metabolic engineering provides insight into the regulation of thiamin biosynthesis in plants

Abstract Thiamin (or thiamine) is a water-soluble B-vitamin (B1), which is required, in the form of thiamin pyrophosphate (TPP), as an essential cofactor in crucial carbon metabolism reactions in all forms of life. To ensure adequate metabolic functioning, humans rely on a sufficient dietary supply of thiamin. Increasing thiamin levels in plants via metabolic engineering is a powerful strategy to alleviate vitamin B1 malnutrition and thus improve global human health. These engineering strategies rely on comprehensive knowledge of plant thiamin metabolism and its regulation. Here, multiple metabolic engineering strategies were examined in the model plant Arabidopsis thaliana. This was achieved by constitutive overexpression of the three biosynthesis genes responsible for B1 synthesis, HMP-P synthase (THIC), HET-P synthase (THI1) and HMP-P kinase/TMP pyrophosphorylase (TH1), either separate or in combination. By monitoring the levels of thiamin, its phosphorylated entities, and its biosynthetic intermediates, we gained insight into the effect of either strategy on thiamin biosynthesis. Moreover, expression analysis of thiamin biosynthesis genes showed the plant’s intriguing ability to respond to alterations in the pathway. Overall, we revealed the necessity to balance the pyrimidine and thiazole branches of thiamin biosynthesis and assessed its biosynthetic intermediates. Furthermore, the accumulation of non-phosphorylated intermediates demonstrated the inefficiency of endogenous thiamin salvage mechanisms. These results serve as guidelines in the development of novel thiamin metabolic engineering strategies.

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Developmental maturation of the colonic uptake process of the microbiota-generated thiamin pyrophosphate

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00067.2021 ◽

2021 ◽

Author(s):

Subrata Sabui ◽

Jose M Romero ◽

Hamid M Said

Keyword(s):

Developmental Regulation ◽

Cell Metabolism ◽

Active Form ◽

Vitamin B1 ◽

Water Soluble ◽

Biologically Active ◽

Nuclear Factors ◽

Water Soluble Vitamin ◽

Uptake Process ◽

Thiamin Pyrophosphate

The water-soluble vitamin B1 is essential for normal human health and physiology. In its main biologically active form, i.e., thiamin pyrophosphate (TPP), the vitamin plays many critical roles in cell metabolism; thus, its deficiency leads to a variety of adverse effects. Humans/mammals obtain vitamin B1 from two exogenous sources: diet and gut microbiota. Considerable amount of the microbiota-generated vitamin B1 exists in the form of TPP, and colonocytes can efficiently absorb this TPP via a high-affinity and specific carrier-mediated mechanism that involves the recently cloned colonic TPP transporter (cTPPT; product of SLC44A4 gene). There is nothing currently known about colonic uptake of TPP during early stages of life, and whether the process undergoes developmental regulation. We addressed this issue using the mouse as animal model. Our results showed that colonic uptake of TPP undergoes developmental up-regulation as the animal moves from the suckling period to weanling and adulthood. This up-regulation in uptake was found to be associated with a parallel induction in level of expression of the cTPPT protein, mRNA and heterologous nuclear RNA (hnRNA), suggesting possible involvement of transcriptional mechanism(s). We also found a parallel up-regulation in level of expression of the two nuclear factors that drive activity of the SLC44A4 promoter (i. e., CREB-1 and Elf-3) with maturation. These results demonstrate, for the first time, that colonic TPP uptake process and cTPPT expression are developmentally up-regulated, and that this up-regulation is likely driven via transcriptional mechanism(s).

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Thiamine (Vitamin B1)

Journal of Evidence-Based Complementary & Alternative Medicine ◽

10.1177/1533210110392941 ◽

2011 ◽

Vol 16 (1) ◽

pp. 12-20 ◽

Cited By ~ 28

Author(s):

Aviva Fattal-Valevski

Keyword(s):

Cognitive Impairment ◽

Cerebral Metabolism ◽

Oxidant Stress ◽

Vitamin B1 ◽

Vitamin B ◽

Deficient Diet ◽

Common Cause ◽

Korsakoff Syndrome ◽

Reducing Substances ◽

B Vitamin

Thiamine (vitamin B1) was the first B vitamin to have been identified. It serves as a cofactor for several enzymes involved in energy metabolism. The thiamine-dependent enzymes are important for the biosynthesis of neurotransmitters and for the production of reducing substances used in oxidant stress defenses, as well as for the synthesis of pentoses used as nucleic acid precursors. Thiamine plays a central role in cerebral metabolism. Its deficiency results in dry beriberi, a peripheral neuropathy, wet beriberi, a cardiomyopathy with edema and lactic acidosis, and Wernicke—Korsakoff syndrome, whose manifestations consist of nystagmus, ophthalmoplegia, and ataxia evolving into confusion, retrograde amnesia, cognitive impairment, and confabulation. Patients on a strict thiamine-deficient diet display a state of severe depletion within 18 days. The most common cause of thiamine deficiency in affluent countries is either alcoholism or malnutrition in nonalcoholic patients. Treatment by thiamine supplementation is beneficial for diagnostic and therapeutic purposes.

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Technical and Economic Feasibility of a Stable Yellow Natural Colorant Production from Waste Lemon Peel

10.20944/preprints202009.0232.v1 ◽

2020 ◽

Author(s):

Rosaria Ciriminna ◽

Billy Forest ◽

Francesco Meneguzzo ◽

Mario Pagliaro ◽

Mark Hamann

Keyword(s):

Physical Stability ◽

Economic Feasibility ◽

Water Soluble ◽

Hydrodynamic Cavitation ◽

Natural Colorant ◽

Lemon Peel ◽

Citrus Flavonoids ◽

Extraction Processes ◽

Solid Liquid ◽

Insight Into

A brief technical and economic insight into producing the water-soluble yellow colorant limocitrol 3-O-6”-[3-hydroxyl-3-methylglutaryl)])-β-D-glucopyranoside from waste lemon peel via simple solid-liquid extraction in aqueous ethanol or via hydrodynamic cavitation of waste lemon peel in water, shows that the biocolorant can be obtained at affordable cost. Coupled to the simplicity and sustainability of the extraction processes suggested, the high chemical and physical stability of this polymethoxylated flavanol and the health benefits of citrus flavonoids, support industrialization of this new bioeconomy production.

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Probing the promiscuity of ent-kaurene oxidases via combinatorial biosynthesis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1512096113 ◽

2016 ◽

Vol 113 (9) ◽

pp. 2526-2531 ◽

Cited By ~ 27

Author(s):

Sibongile Mafu ◽

Meirong Jia ◽

Jiachen Zi ◽

Dana Morrone ◽

Yisheng Wu ◽

...

Keyword(s):

Natural Products ◽

Metabolic Engineering ◽

Substrate Specificity ◽

Plant Hormone ◽

Structural Similarity ◽

Combinatorial Biosynthesis ◽

Underlying Structure ◽

Engineering System ◽

Insight Into

The substrate specificity of enzymes from natural products’ metabolism is a topic of considerable interest, with potential biotechnological use implicit in the discovery of promiscuous enzymes. However, such studies are often limited by the availability of substrates and authentic standards for identification of the resulting products. Here, a modular metabolic engineering system is used in a combinatorial biosynthetic approach toward alleviating this restriction. In particular, for studies of the multiply reactive cytochrome P450, ent-kaurene oxidase (KO), which is involved in production of the diterpenoid plant hormone gibberellin. Many, but not all, plants make a variety of related diterpenes, whose structural similarity to ent-kaurene makes them potential substrates for KO. Use of combinatorial biosynthesis enabled analysis of more than 20 such potential substrates, as well as structural characterization of 12 resulting unknown products, providing some insight into the underlying structure–function relationships. These results highlight the utility of this approach for investigating the substrate specificity of enzymes from complex natural products’ biosynthesis.

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INFLUENCE OF TAP WATER CONTAMINANTS ON THE COMPONENTS OF REDUCED WHEY

Известия вузов. Пищевая технология ◽

10.26297/0579-3009.2020.4.5 ◽

2020 ◽

Author(s):

А.К. ГОРЕЛКИНА ◽

И.В. ТИМОЩУК ◽

Н.С. БАГДОНАС

Keyword(s):

Vitamin C ◽

Organic Contaminants ◽

Vitamin B6 ◽

Vitamin B1 ◽

Water Soluble ◽

Gas Liquid Chromatography ◽

Vitamin B2 ◽

Water Contaminants ◽

Whey Beverages ◽

Control Samples

Изучена стойкость нутриентов молокосырья – белков, лактозы, витаминов С и группы В, используемого в производстве сывороточных напитков в присутствии приоритетных органических контаминантов – трихлорэтилена, хлороформа и дихлорэтана, которые обладают токсическим и канцерогенным действием и образуются при хлорировании в процессе водоподготовки в воде, применяемой для производства восстановленных и рекомбинированных молочных продуктов. Содержание белков, лактозы в восстановленной сыворотке определяли методом рефрактометрии; водорастворимых витаминов – методом капиллярного электрофореза; хлороформа, трихлорэтилена и дихлорэтана – методом газожидкостной хроматографии. Установлено, что хлороформ в воде не оказывает влияния на сохранность лактозы, белков и витаминов при приготовлении восстановленной сыворотки. Отмечено снижение содержания белков сыворотки, приготовленной на воде в присутствии трихлорэтилена и дихлорэтана, на 11%, лактозы – на 32% в сравнении с контрольными образцами, произведенными на воде без органических контаминантов. Содержание витаминов в восстановленной молочной сыворотке в присутствии трихлорэтилена снизилось: С – на 19%, В1 – на 28%, В2 – на 53%, В6 – на 8%; для дихлорэтана содержание витаминов снизилось: С – на 17%, В1 – на 36%, В2 – на 38%, В6 – на 36% в сравнении с контрольными образцами без органических примесей. Теоретически обоснован механизм взаимодействия белков, лактозы, витаминов восстановленной сыворотки с трихлорэтиленом и дихлорэтаном. Предложено для предотвращения снижения качества готового продукта воду, используемую для производства сывороточных напитков, подвергать дополнительной очистке от галогенорганических контаминантов. The stability of milk raw materials’ nutrients – proteins, lactose, vitamins C and B used in the production of whey beverages in the presence of priority organic contaminants – trichloroethylene, chloroform and dichloroethane, which have a toxic and carcinogenic effect and are formed during chlorination during water treatment in water used for the production of reduced and recombined dairy products was studied. The content of proteins and lactose in the reduced serum was determined by refractometry; water-soluble vitamins – by capillary electrophoresis; chloroform, trichloroethylene and dichloroethane – by gas-liquid chromatography. It was found that chloroform in water does not affect the safety of lactose, proteins and vitamins in the preparation of reduced whey. There was a decrease in the content of serum proteins prepared in water in the presence of trichloroethylene and dichloroethane by 11%, and lactose – by 32% in comparison with control samples produced in water without organic contaminants. Vitamin C content in the recovered whey in the presence of trichloroethylene has declined by 19%, vitamin B1 – 28%, vitamin B2 – 53%, vitamin B6 – 8%; for dichloroethane content of vitamin C decreased by 17%, vitamin B1 – 36%, vitamin B2 – 38%, vitamin B6 – 36% in comparison with control samples without organic impurity. The mechanism of interaction of proteins, lactose, and reduced serum vitamins with trichloroethylene and dichloroethane is theoretically justified. Therefore, to prevent a decrease in the quality of the finished product, the water used for the production of whey beverages must first be subjected to additional purification from organohalogen contaminants.

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Ultradeformable vesicles: concepts and applications relating to the delivery of skin cosmetics

Therapeutic Delivery ◽

10.4155/tde-2021-0044 ◽

2021 ◽

Author(s):

Andang Miatmoko ◽

Qurrota Ayunin ◽

Widji Soeratri

Keyword(s):

Skin Aging ◽

Water Soluble ◽

In Vivo Studies ◽

Active Ingredients ◽

Self Confidence ◽

Skin Delivery ◽

Cosmetic Ingredients ◽

Insight Into

Skin aging is a phenomenon resulting in reduced self-confidence, thus becoming a major factor in social determinants of health. The use of active cosmetic ingredients can help prevent skin aging. Transfersomes are well known to be capable of deeply penetrating the dermis. This scoping review provides an insight into transfersomes and their prospective use in anti-aging cosmetics. Numerous reports exist highlighting the successful skin delivery of therapeutic agents such as high-molecular-weight, poorly water soluble and poorly permeable active ingredients by means of transfersomes. Moreover, in vitro and in vivo studies have indicated that transfersomes increase the deposition, penetration and efficacy of active ingredients. However, the use of transfersomes in the delivery of active cosmetic ingredients is limited. Considering their similar physicochemical properties, transfersomes should possess considerable potential as a delivery system for anti-aging cosmetics.

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Riboflavin: The Health Benefits of a Forgotten Natural Vitamin

International Journal of Molecular Sciences ◽

10.3390/ijms21030950 ◽

2020 ◽

Vol 21 (3) ◽

pp. 950 ◽

Cited By ~ 12

Author(s):

Nittiya Suwannasom ◽

Ijad Kao ◽

Axel Pruß ◽

Radostina Georgieva ◽

Hans Bäumler

Keyword(s):

Protective Effect ◽

Toxic Effect ◽

Biological Effects ◽

Health Benefits ◽

Water Soluble ◽

Medical Conditions ◽

Protective Properties ◽

Anti Cancer ◽

Anti Oxidant ◽

B Vitamin

Riboflavin (RF) is a water-soluble member of the B-vitamin family. Sufficient dietary and supplemental RF intake appears to have a protective effect on various medical conditions such as sepsis, ischemia etc., while it also contributes to the reduction in the risk of some forms of cancer in humans. These biological effects of RF have been widely studied for their anti-oxidant, anti-aging, anti-inflammatory, anti-nociceptive and anti-cancer properties. Moreover, the combination of RF and other compounds or drugs can have a wide variety of effects and protective properties, and diminish the toxic effect of drugs in several treatments. Research has been done in order to review the latest findings about the link between RF and different clinical aberrations. Since further studies have been published in this field, it is appropriate to consider a re-evaluation of the importance of RF in terms of its beneficial properties.

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An Improved Method for the Quantification of Thiamine (Vitamin B1) in Cocoa-Containing Beverage Powders

Journal of AOAC International ◽

10.5740/jaoacint.17-0012 ◽

2017 ◽

Vol 100 (5) ◽

pp. 1511-1515

Author(s):

Frédéric Martin ◽

Liliane Meyer ◽

Konstantinos Zelianos ◽

Esther Campos Gimenez

Keyword(s):

Ph Value ◽

Solid Phase ◽

Vitamin B1 ◽

Water Soluble ◽

Local Market ◽

Improved Method ◽

Test Portion ◽

Beverage Samples ◽

Water Soluble Vitamin ◽

Thiamine Content

Abstract The purpose of this work was to understand low recoveries of thiamine (vitamin B1) when extracted from cocoa-containing beverage powders fortified with water-soluble vitamin B1, and to develop and validate a new procedure to improve these results. Based on the literature, previous trials have focused on two main factors: pH value prior to paper filtration and the need for solid-phase extraction (SPE) clean up. We demonstrate that by following European Standard EN 14122, recovery of thiaminein cocoa-containing beverage powders is low and dependent on the test portion (86 and 72% for 0.5 and 1.5 g test portions, respectively). Our improved method resolved this problem by keeping the pH low (around 1) prior to paper filtration, leading to a 96.3% recovery and high precision (RSDr of 3.5%). The use of strong cation-exchange SPE cartridges for cleanup prior to the thiamine oxidation reaction proved to be essential. A comparison between our improved method and EN 14122 on nine cocoa-containing beverage samples available on local market from different manufacturers showed a systematic increase in thiamine content (up to 70%) when the improved methodwas applied. The highest difference was observed forthe sample that contained the highest amount of cocoa. However, for beverage powders that contained bothcocoa and milk, no difference was observed.

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Insight into the self-assembly of water-soluble perylene bisimide derivatives through a combined computational and experimental approach

Nanoscale ◽

10.1039/c9nr03898a ◽

2019 ◽

Vol 11 (34) ◽

pp. 15917-15928 ◽

Cited By ~ 5

Author(s):

Emily R. Draper ◽

Liam Wilbraham ◽

Dave J. Adams ◽

Matthew Wallace ◽

Ralf Schweins ◽

...

Keyword(s):

Amino Acid ◽

Self Assembly ◽

Experimental Approach ◽

Water Soluble ◽

The Self ◽

Experimental Techniques ◽

Perylene Bisimide ◽

Perylene Bisimides ◽

Insight Into

We use a combination of computational and experimental techniques to study the self-assembly and gelation of amino-acid functionalised water-soluble perylene bisimides.

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