hydroxypropionic acid
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Author(s):  
Zhishuai Chang ◽  
Wei Dai ◽  
Yufeng Mao ◽  
Zhenzhen Cui ◽  
Zhidan Zhang ◽  
...  

Acetate is an economical and environmental-friendly alternative carbon source. Herein, the potential of harnessing Corynebacterium glutamicum as a host to produce 3-hydroxypropionic acid (3-HP) from acetate was explored. First, the expression level of malonyl-CoA reductase from Chloroflexus aurantiacus was optimized through several strategies, strain Cgz2/sod-N-C* showed an MCR enzyme activity of 63 nmol/mg/min and a 3-HP titer of 0.66 g/L in flasks. Next, the expression of citrate synthase in Cgz2/sod-N-C* was weakened to reduce the acetyl-CoA consumption in the TCA cycle, and the resulting strain Cgz12/sod-N-C* produced 2.39 g/L 3-HP from 9.32 g/L acetate. However, the subsequent deregulation of the expression of acetyl-CoA carboxylase genes in Cgz12/sod-N-C* resulted in an increased accumulation of intracellular fatty acids, instead of 3-HP. Accordingly, cerulenin was used to inhibit fatty acid synthesis in Cgz14/sod-N-C*, and its 3-HP titer was further increased to 4.26 g/L, with a yield of 0.50 g 3-HP/g-acetate. Finally, the engineered strain accumulated 17.1 g/L 3-HP in a bioreactor without cerulenin addition, representing the highest titer achieved using acetate as substrate. The results demonstrated that Corynebacterium glutamicum is a promising host for 3-HP production from acetate.


Author(s):  
Sarang S. Bhagwat ◽  
Yalin Li ◽  
Yoel R. Cortés-Peña ◽  
Emma C. Brace ◽  
Teresa A. Martin ◽  
...  

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Florence de Fouchécour ◽  
Anaïs Lemarchand ◽  
Henry-Éric Spinnler ◽  
Claire Saulou-Bérion

AbstractAcetic acid bacteria (AAB) can selectively oxidize diols into their corresponding hydroxyacids. Notably, they can convert 1,3-propanediol (1,3-PDO) into 3-hydroxypropionic acid (3-HP), which is a promising building-block. Until now, 3-HP production with AAB is carried out in batch and using resting cells at high cell densities (up to 10 g L−1 of cell dry weight). This approach is likely limited by detrimental accumulation of the intermediate 3-hydroxypropanal (3-HPA). Herein, we investigate an alternative implementation that allows highly efficient 3-HP production with lower cell densities of growing cells and that prevents 3-HPA accumulation. First, growth and 3-HP production of Acetobacter sp. CIP 58.66 were characterized with 1,3-PDO or glycerol as growth substrate. The strain was then implemented in a bioreactor, during a sequential process where it was first cultivated on glycerol, then the precursor 1,3-PDO was continuously supplied at a varying rate, easily controlled by the pH control. Different pH set points were tested (5.0, 4.5, and 4.0). This approach used the natural resistance of acetic acid bacteria to acidic conditions. Surprisingly, when pH was controlled at 5.0, the performances achieved in terms of titer (69.76 g3-HP L−1), mean productivity (2.80 g3-HP L−1 h−1), and molar yield (1.02 mol3-HP mol−11,3-PDO) were comparable to results obtained with genetically improved strains at neutral pH. The present results were obtained with comparatively lower cell densities (from 0.88 to 2.08 g L−1) than previously reported. This feeding strategy could be well-suited for future scale-up, since lower cell densities imply lower process costs and energy needs.


2021 ◽  
Author(s):  
Florence DE FOUCHÉCOUR ◽  
Anaïs LEMARCHAND ◽  
Henry-Éric SPINNLER ◽  
Claire SAULOU-BÉRION

Abstract Acetic acid bacteria (AAB) can selectively oxidize diols into their corresponding hydroxyacids. Notably, they can convert 1,3-propanediol (1,3-PDO) into 3-hydroxypropionic acid (3-HP), which is a promising building-block. Until now, 3-HP production with AAB is carried out in batch and using resting cells at high cell densities (up to 10 g L− 1 of cell dry weight). This approach is likely limited by detrimental accumulation of the intermediate 3-hydroxypropanal (3-HPA). Herein, we investigate an alternative implementation that allows highly efficient 3-HP production with lower cell densities of growing cells and that prevents 3-HPA accumulation. First, growth and 3-HP production of Acetobacter sp. CIP 58.66 were characterized with 1,3-PDO or glycerol as growth substrate. The strain was then implemented in a bioreactor, during a sequential process where it was first cultivated on glycerol, then the precursor 1,3-PDO was continuously supplied at a varying rate, easily controlled by the pH control. Different pH set points were tested (5.0, 4.5, and 4.0). This approach used the natural resistance of acetic acid bacteria to acidic conditions. Surprisingly, when pH was controlled at 5.0, the performances achieved in terms of titer (69.76 g3 − HP L− 1), mean productivity (2.80 g3 − HP L− 1 h− 1), and molar yield (1.02 mol3 − HP mol− 11,3−PDO) were comparable to results obtained with genetically improved strains at neutral pH. The present results were obtained with comparatively lower cell densities (from 0.88 to 2.08 g L− 1) than previously reported. This feeding strategy could be well-suited for future scale-up, since lower cell densities imply lower process costs and energy needs.


Author(s):  
А.С. Галушкин ◽  
Е.Ю. Пыркова ◽  
М.В. Куркина ◽  
С.В. Михайлова ◽  
В.П. Воронцова ◽  
...  

Пропионовая ацидемия (ПА) - редкое наследственное заболевание с аутосомно-рецессивным типом наследования, относится к «классическим», т.е. наиболее часто диагностируемым органическим ацидемиям. В данной работе представлена биохимическая и молекулярно-генетическая характеристика 16 пациентов с ПА, выявленных в Российской Федерации. У большинства обследованных пациентов первые симптомы проявились в первые месяцы жизни, наиболее частые из них: нарушения вскармливания, судороги, мышечная гипотония, угнетение сознания. При биохимическом исследовании (ГХ-МС и МС/МС) у пациентов было выявлено повышение концентрации 3-гидроксипропионой кислоты, метилцитрата, пропионилглицина, C3, глицина, которые являются патогномоничными маркерами этой патологии. В результате молекулярно-генетического анализа в генах PCCA и PCCB было выявлено 6 неописанных ранее вариантов нуклеотидной последовательности (4 варианта в гене PCCA, 2 - в гене PCCB), один из которых (PCCB: c.655-2A>G) имеет высокую частоту (17,2%, 5/29 мутантных аллелей генов PCCA и PCCB) в исследуемой выборке. Локализация миссенс-вариантов и их влияние на структуру белка были продемонстрированы на 3D-модели фермента пропионил-КоА-карбоксилазы. Выявленным вариантам были присвоены критерии патогенности в соответствии с рекомендациями American College of Medical Genetics and Genomics (ACMG). Propionic acidemia is a rare autosomal recessive metabolic disorder, characterized as classic organic acidemia. The article represents biochemical and molecular characterization of 16 patients diagnosed with propionic acidemia in Russia. Symptoms appeared during the first months after birth in most cases. Poor feeding, seizures, hypotonia, lethargy were the most frequent symptoms. Biochemical tests (GC-MS and MS/MS) showed elevated 3-hydroxypropionic acid, methylcitrate, propionylglycine, C3 in patients blood. Six undescribed earlier variants were found as a result of PCCA & PCCB (4 variants - PCCA, 2 - PCCB) genes DNA-tests. Variant c.655-2A>G in PCCB is the most frequent in this group (17.2%). Localization of missense variants and their effect on protein was demonstrated using propionyl-CoA carboxylase 3D model. New variants were classified according to the American College of Medical Genetics and Genomics (ACMG) guidelines.


Author(s):  
Albert Fina ◽  
Gabriela Coelho Brêda ◽  
Míriam Pérez‐Trujillo ◽  
Denise Maria Guimarães Freire ◽  
Rodrigo Volcan Almeida ◽  
...  

2021 ◽  
Vol 329 ◽  
pp. 124867
Author(s):  
Trinh Thi Nguyen ◽  
Suman Lama ◽  
Satish Kumar Ainala ◽  
Mugesh Sankaranarayanan ◽  
Ashish Singh Chauhan ◽  
...  

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