scholarly journals Challenges and opportunities of bioprocessing 5-aminolevulinic acid using genetic and metabolic engineering: a critical review

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Ying-Chen Yi ◽  
I-Tai Shih ◽  
Tzu-Hsuan Yu ◽  
Yen-Ju Lee ◽  
I-Son Ng

Abstract5-Aminolevulinic acid (5-ALA), a non-proteinogenic five-carbon amino acid, has received intensive attentions in medicine due to its approval by the US Food and Drug Administration (FDA) for cancer diagnosis and treatment as photodynamic therapy. As chemical synthesis of 5-ALA performed low yield, complicated processes, and high cost, biosynthesis of 5-ALA via C4 (also called Shemin pathway) and C5 pathway related to heme biosynthesis in microorganism equipped more advantages. In C4 pathway, 5-ALA is derived from condensation of succinyl-CoA and glycine by 5-aminolevulic acid synthase (ALAS) with pyridoxal phosphate (PLP) as co-factor in one-step biotransformation. The C5 pathway involves three enzymes comprising glutamyl-tRNA synthetase (GltX), glutamyl-tRNA reductase (HemA), and glutamate-1-semialdehyde aminotransferase (HemL) from α-ketoglutarate in TCA cycle to 5-ALA and heme. In this review, we describe the recent results of 5-ALA production from different genes and microorganisms via genetic and metabolic engineering approaches. The regulation of different chassis is fine-tuned by applying synthetic biology and boosts 5-ALA production eventually. The purification process, challenges, and opportunities of 5-ALA for industrial applications are also summarized.

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Ying-Chen Yi ◽  
I-Son Ng

AbstractProgramming non-canonical organisms is more attractive due to the prospect of high-value chemical production. Among all, Shewanella oneidensis MR-1 possesses outstanding heme synthesis ability and is well-known for electron transfer, thus has high potential in microbial fuel cell and bioremediation. However, heme, as the final product of C4 and C5 pathways, is regulated by heme cluster for the high-value 5-aminolevulinic acid (ALA) for cancer photodynamic therapy, which has never been explored in MR-1. Herein, the heme metabolism in MR-1 was firstly optimized for ALA production. We applied CRISPR interference (CRISPRi) targeted on the genes to fine-tune carbon flux in TCA cycle and redirected the carbon out-flux from heme, leading to a significant change in the amino acid profiles, while downregulation of the essential hemB showed a 2-fold increasing ALA production via the C5 pathway. In contrast, the modular design including of glucokinase, GroELS chaperone, and ALA synthase from Rhodobacter capsulatus enhanced ALA production markedly in the C4 pathway. By integrating gene cluster under dual T7 promoters, we obtained a new strain M::TRG, which significantly improved ALA production by 145-fold. We rewired the metabolic flux of MR-1 through this modular design and successfully produced the high-value ALA compound at the first time.


2021 ◽  
Vol 22 (9) ◽  
pp. 4949
Author(s):  
Veronika Kazimírová ◽  
Martin Rebroš

The production of aldehydes, highly reactive and toxic chemicals, brings specific challenges to biocatalytic processes. Absence of natural accumulation of aldehydes in microorganisms has led to a combination of in vitro and in vivo strategies for both, bulk and fine production. Advances in genetic and metabolic engineering and implementation of computational techniques led to the production of various enzymes with special requirements. Cofactor synthesis, post-translational modifications and structure engineering are applied to prepare active enzymes for one-step or cascade reactions. This review presents the highlights in biocatalytical production of aldehydes with the potential to shape future industrial applications.


2020 ◽  
Vol 1008 ◽  
pp. 33-38
Author(s):  
Marwa Nabil ◽  
Hussien A. Motaweh

Silica is one of the most important materials used in many industries. The basic factor on which the selection process depends is the structural form, which is dependent on the various physical and chemical properties. One of the common methods in preparing pure silica is that it needs more than one stage to ensure the preparation process completion. The goal of this research is studying the nucleation technique (Bottom-top) for micro-wires and micro-ribbons silica synthesis. The silica nanoand microstructures are prepared using a duality (one step); a combination of alkali chemical etching process {potassium hydroxide (3 wt %) and n-propanol (30 Vol %)} and the ultra-sonication technique. In addition, the used materials in the preparation process are environmentally friendly materials that produce no harmful residues. The powder product is characterized using XRD, FTIR, Raman spectrum and SEM for determining the shape of architectures. The most significant factor of the nucleation mechanism is the sonication time of silica powder production during the dual technique. The product stages are as follows; silica nanoparticles (21-38 nm), nanoclusters silica (46 – 67 nm), micro-wires silica (1.17 – 6.29 μm), and micro-ribbons silica (19.4 – 54.1 μm). It's allowing for use in environmental applications (multiple wastewater purification, multiple uses in air filters, as well as many industrial applications).


2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Zhenyu Zhang ◽  
Pengfu Liu ◽  
Weike Su ◽  
Huawei Zhang ◽  
Wenqian Xu ◽  
...  

AbstractTrans-4-hydroxy-l-proline is an important amino acid that is widely used in medicinal and industrial applications, particularly as a valuable chiral building block for the organic synthesis of pharmaceuticals. Traditionally, trans-4-hydroxy-l-proline is produced by the acidic hydrolysis of collagen, but this process has serious drawbacks, such as low productivity, a complex process and heavy environmental pollution. Presently, trans-4-hydroxy-l-proline is mainly produced via fermentative production by microorganisms. Some recently published advances in metabolic engineering have been used to effectively construct microbial cell factories that have improved the trans-4-hydroxy-l-proline biosynthetic pathway. To probe the potential of microorganisms for trans-4-hydroxy-l-proline production, new strategies and tools must be proposed. In this review, we provide a comprehensive understanding of trans-4-hydroxy-l-proline, including its biosynthetic pathway, proline hydroxylases and production by metabolic engineering, with a focus on improving its production.


2021 ◽  
Author(s):  
Valentina Donadei ◽  
Heli Koivuluoto ◽  
Essi Sarlin ◽  
Petri Vuoristo

Abstract In subzero conditions, atmospheric ice naturally accretes on surfaces in outdoor environments. This accretion can compromise the operational performance of several industrial applications, such as wind turbines, power lines, aviation, and maritime transport. To effectively prevent icing problems, the development of durable icephobic coating solutions is strongly needed. Here, the durability of lubricated icephobic coatings was studied under repeated icing/deicing cycles. Lubricated coatings were produced in one-step by flame spraying with hybrid feedstock injection. The coating icephobicity was investigated by accreting ice from supercooled microdroplets using an icing wind tunnel. The ice adhesion strength was evaluated by a centrifugal ice adhesion tester. The icing performance was investigated over four icing/deicing cycles. Surface properties of coatings, such as morphology, topography, chemical composition and wettability, were analyzed before and after the cycles. The results showed an increase in ice adhesion over the cycles, while a stable icephobic behaviour was retained for one selected coating. Moreover, consecutive ice detachment caused a surface roughness increase. This promotes the formation of mechanical interlocking with ice, thus justifying the increased ice adhesion. Finally, the coating hydrophobicity mainly decreased as a consequence of the damaged surface topography. In summary, lubricated coatings retained a good icephobic level after the cycles, thus demonstrating their potential for icephobic applications.


2021 ◽  
Vol 10 (1) ◽  
pp. 39
Author(s):  
Xinhua Qi ◽  
Wenlong Yan ◽  
Zhibei Cao ◽  
Mingzhu Ding ◽  
Yingjin Yuan

Polyethylene terephthalate (PET) is a widely used plastic that is polymerized by terephthalic acid (TPA) and ethylene glycol (EG). In recent years, PET biodegradation and bioconversion have become important in solving environmental plastic pollution. More and more PET hydrolases have been discovered and modified, which mainly act on and degrade the ester bond of PET. The monomers, TPA and EG, can be further utilized by microorganisms, entering the tricarboxylic acid cycle (TCA cycle) or being converted into high value chemicals, and finally realizing the biodegradation and bioconversion of PET. Based on synthetic biology and metabolic engineering strategies, this review summarizes the current advances in the modified PET hydrolases, engineered microbial chassis in degrading PET, bioconversion pathways of PET monomers, and artificial microbial consortia in PET biodegradation and bioconversion. Artificial microbial consortium provides novel ideas for the biodegradation and bioconversion of PET or other complex polymers. It is helpful to realize the one-step bioconversion of PET into high value chemicals.


2019 ◽  
Author(s):  
Yingshuo Hou ◽  
Siyu Chen ◽  
Jianjun Wang ◽  
Guizhen Liu ◽  
Sheng Wu ◽  
...  

Abstract Background:Corynebacterium ammoniagenes is an important industrial organism that is widely used to produce nucleotides and the potential for industrial production of coenzyme A by Corynebacterium ammoniagenes ATCC 6871 has been shown. However, the yield of coenzyme A needs to be improved and the available constitutive promoters are rather limited in this strain. Results:In this study, 20 putative DNA promoters derived from genes with high transcription levels and 6 promoters from molecular chaperone genes were identified. To evaluate the activity of each promoter, red fluorescence protein (RFP) was used as a reporter. We successfully isolated a range of promoters with different activity levels resulting in different fluorescent intensities. A fragment derived from the upstream sequence of the 50S ribosomal protein L21 (Prpl21) exhibited the strongest activity among the 26 identified promoters. Furthermore, type III pantothenate kinase from Pseudomonas putida (PpcoaA) was overexpressed in C. ammoniagenes under the control of Prpl21, CoA yield increased approximately 4.1 times. Conclusions:This study provides a paradigm for rational isolation of promoters with different activities and their application in metabolic engineering. These promoters will enrich the available promoter toolkit for C. ammoniagenes and should be valuable in current platforms for metabolic engineering and synthetic biology for the optimization of pathways to extend the product spectrum or improve the productivity in C. ammoniagenes ATCC 6871 for industrial applications.


2021 ◽  
Author(s):  
Bashar Amer ◽  
Ramu Kakumanu ◽  
yangtian not provided ◽  
Aymerick Eudes ◽  
Edward EK Baidoo

Cell-wall-bound (CWB) aromatics such as ferulate and p-coumarate play important physiological roles in plant development and response to stresses. Their presence also poses some challenges and opportunities during processing of plant biomass in various agro-industrial applications. To this end, we have developed a robust high-throughput reversed-phase liquid chromatography mass spectrometry method for quantifying CWB phenolic compounds. The method showed excellent linearity (R2 = ≥0.999) and intraday retention time repeatability (≤ 0.31 %RSD) for ferulate and p-coumarate. The limits of detection and quantitation for these analytes were ≤ 39 nM and 130 nM, respectively. Furthermore, there was very little effect of the CWB sample matrix on the retention times of the analytes and analyte percent recoveries from the CWB sample matrix was ≥83.91%.


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