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In this work, acyltransferase activity of a new bacterial isolate Bacillus smithii strain IITR6b2 was utilized for the synthesis of nicotinic acid hydroxamate (NAH), a heterocyclic class of hydroxamic acid. NAH is an important pyridine derivative and has found its role as bioligand, urease inhibitor, antityrosinase, antioxidant, antimetastatic, and...
Contexts in source publication
Context 1
... achieve higher yield of hydroxamic acid by mini- mizing the undesirable by-product (acid) formation from amide, it is desirable to have a biocatalyst with high acyltransferase activity and minimum amide and hydroxamic acid hydrolase activities (Fig. 1). The aim of the present study is to develop a process for synthesis of nicotinic acid hydroxamate from nicotinamide using acyl- transferase activity of Bacillus smithii strain IITR6b2. In this work, special emphasis was placed on process devel- opment for NAH production with minimum formation of the by-product nicotinic acid. Different ...
Context 2
... of molar extinction coefficient for nicotinic acid hydroxamate/Fe(III) complex Molar extinction coefficient ([ M ) of the NAH/Fe(III) complex was determined to quantify NAH formed in the acyltransferase reaction. The [ M value of 4.14 9 10 2 l mol -1 cm -1 was obtained from the slope of a standard curve (Supplementary Fig. S1). Fournand et al. [7] determined the [ M values of aliphatic saturated, aliphatic unsaturated and a, b, c-amino hydroxamic acids but this is the first report on determination of molar extinction coefficient ([ M ) for heterocyclic class of hydroxamic acids. ...
Citations
... Enzyme inhibition is a crucial factor in biocatalysis reaction, therefore, to avoid substrate inhibition, a fed-batch process was performed. After two feeding (200 mM at 40 min), 89.4 % of bioconversion was achieved with 58.5 wt % nicotinic acid hydroxamate (95 % purity) recovery yield [34]. ...
In recent years, biocatalysts have emerged as crucial tool in organic synthesis, particularly for the production of drug intermediates and precursors, e.g., the synthesis of hydroxamic acids. Traditionally, hydroxamic acids were synthesized using organic chemistry methods. However, with the growing emphasis on sustainable and environment‐friendly practices, the chemical industry has increasingly turned towards green synthesis approaches. The significance of hydroxamic acids in medicinal chemistry has also contributed to the changing trends. Following the approval of certain hydroxamic acids as histone deacetylase (HDAC) inhibitors for cancer treatment by the Food and Drug Administration (US‐FDA), there has been a renewed focus on their synthesis and the development of derivatives with improved properties. As an alternative route, amidases have emerged as promising biocatalysts for hydroxamic acid synthesis through their acyltransferase activity. Recent advancements in the synthesis approaches for hydroxamic acids are reviewed. The biocatalytic routes are explored, emphasizing the use of amidases and their acyltransferase activity. The scope and potential applications of this chemoenzymatic approach in synthesizing various hydroxamic acids and their derivatives are discussed. Such advancements have the potential to revolutionize the production of these important compounds, making the synthesis process more sustainable, efficient, and economically viable.
... Other reported amidases i.e. Bacillus smithii strain IITR6b2 (Agarwal et al. 2013), and Rhodococcus pyridinivorans NIT-36 (Ansu et al. 2016) exhibited maximum acyltransferase activity around neutral pH. ...
... MTCC 10674 was reported as 50ºC with maximum acyltransferase activity (Bhatiya et al. 2013). Further, Bacillus smithii IITR6b2 exhibited maximum biotransformation of nicotinamide into nicotinic acid hydroxamate at 55ºC (Agarwal et al. 2013). Sharma et al. (2012) MTCC 10674 (Bhatiya et al. 2013). ...
Aim:
Presently, N-hydroxy-N'-phenyl-octanediamide (vorinostat) which is an effective histone deacetylase inhibitor, is being synthesized chemically. Hence, present study aims to developed an eco-friendly approach for the synthesis of vorinostat from N'-phenyloctanediamide through biotransformation.
Methods and results:
Using the amidase of Bacillus smithii IIIMB2907 in time course conversion and organic solvent compatibility, maximum bioconversion was observed at 12 hour of reaction time and in presence of ethanol respectively. Potassium phosphate buffer of pH 7.0 supported maximum bioconversion of N'-phenyloctanediamide (10mM) into N-hydroxy-N'- phenyl-octanediamide at 40°C. Bench scale study was successfully carried out with 83% yield of purified vorinostat.
Conclusion:
In this study, an eco-friendly approach for the biotransformation of N'-phenyloctanediamide into vorinostat was developed by using cell free extract of thermophilic strain B. smithii IIIMB2907.
... This is the first report in which acetohydroxamic acid was synthesized using amidase of any theromophilic Bacillus smithii strain. In reported literature, Bacillus smithii strain IITR6b2 was reported for the synthesis of nicotinic acid hydroxamate and isoniazid 13,14 . However, Rhodococcus pyridinivorans NIT-36 was reported for the synthesis of acetohydroxamic acid 15 . ...
Acetohydroxamic acid is a pharmaceutically active metal chelating agent which has various applications in the field of medicine. Current study focuses on the enzymatic synthesis of acetohydroxamic acid catalysed by thermophilic amidase from Bacillus smithii IIIMB2907. Bacterial cells were grown in 7 L fermenter for amidase production and effect of pH, temperature and substrate concentration for the biotransformation of acetamide to acetohydroxamic acid was studied. Batch reaction was also successfully optimized at bench scale with the recovery of ≈ 81% acetohydroxamic acid (purified).
... These enzymes have thus turned out to be attractive biocatalyst for organic synthesis, i.e., carboxylic acids, hydroxamic acids, hydrazides, and also for bioremediation [16]. Their hydrolytic activities are employed in combination with nitrile hydratase for the production of commercially important carboxylic acids [69,96] and amide containing effluent treatment [17], whereas acyltransferase activity is used for the synthesis of hydroxamic acid [18,[97][98][99]. The applications of amidases in the production of various fine chemicals have been reviewed extensively [16]. ...
... Recently, acyl transferase activities of amidases were exploited mainly for the synthesis of pharmaceutically active hydroxamic acids and hydrazides. A number of important hydroxamic acids, i.e., acetohydroxamic acid [98,100], benzohydroxamic acid [20], and nicotinyl hydroxamate [97,99], have been synthesized using acyltransferase activity of amidases (Table 5). Hydroxamic acids (R-CONHOH) are industrially very important as they form chelates with metal ions. ...
... In another study, Bhatia et al. had synthesized 16 g of nicotinyl hydroxamic acid from nicotinamide and hydroxylamine at 1 L scale using free cells of Pseudomonas putida BR1 with 32 g L −1 h −1 volumetric productivity [97]. The Bacillus smithii IITR6b2 exhibiting acyltransferase activity has been explored for the synthesis of another important product, i.e., nicotinic acid hydroxamate [99]. ...
Nitrile metabolizing enzymes, i.e., aldoxime dehydratase, hydroxynitrile lyase, nitrilase, nitrile hydratase, and amidase, are the key catalysts in carbon nitrogen triple bond anabolism and catabolism. Over the past several years, these enzymes have drawn considerable attention as prominent biocatalysts in academia and industries because of their wide applications. Research on various aspects of these biocatalysts, i.e., sources, screening, function, purification, molecular cloning, structure, and mechanisms, has been conducted, and bioprocesses at various scales have been designed for the synthesis of myriads of useful compounds. This review is focused on the potential of nitrile metabolizing enzymes in the production of commercially important fine chemicals such as nitriles, carboxylic acids, and amides. A number of opportunities and challenges of nitrile metabolizing enzymes in bioprocess development for the production of bulk and fine chemicals are discussed.
... The immobilized biocatalyst ex- hibited higher stability and faster reaction rates even at low sub- strate concentrations in comparison to the free form. In some cases, the yields were high up to 80% (Scheme 14) [73] [74]. ...
Background
Hydroxamic acids are a major class of organic compounds. They have a wide variety of pharmacological actions in targeting cancers, cardiovascular diseases, HIV, Alzheimer's disease, Malaria, Allergic diseases.
Objective
The present review focuses on the chemistry of conventional and non-conventional routes for the synthesis of hydroxamic acids reported till date.
Conclusion
The hydroxamic acids are conventionally synthesized via carboxylic acid and their acid chloride derivatives. However, some other functional groups i.e. aldehyde, amine, amide and alcohol can also be converted to hydroxamate with ease. The solid phase synthesis techniques are also gaining importance for the synthesis of hydroxamic acids and these pathways have opened a wide arena for the synthesis of diverse and complex hydroxamic acids.
... A number of traditional approaches are available for HAD preparation by organic synthesis, but some are tedious, time-consuming and costly as well. Bacterial amidases (acrylamide amidohydrolase; EC 3.5.1.4.) can be used for the production of various HAD by catalyzing transamidation reactions using a wide range of amides as substrates and hydroxylamine as acyl group acceptor [21][22][23]. Amidases have significant potential due to their broad substrate specificity and there are numerous reports on the immobilization of microbial cells with amidase activity providing several advantages for its various applications [21,[24][25][26]. ...
In this study were investigated, the synthesis, acetylcholinesterase inhibition and
antioxidant activity of a series of hydroxamic acid derivatives (HAD), with different
chemical group characteristics, such as aliphatic (acetohydroxamic acid and butyryl
hydroxamic acid), aromatic (benzohydroxamic acid and phenylalanine hydroxamic acid)
and amino acid (glycine hydroxamic acid and alanine hydroxamic acid). It was observed
that these HAD compounds present very promising activity as acetylcholinesterase
(AChE) inhibitors and as antioxidants. The aliphatic HAD demonstrated to have a higher
inhibitory activity of AChE than amino acid or aromatic HAD. As for the antioxidant
activity, a high antioxidant potential was found for all the compounds with EC50 values
ranging from 0.19 μM to 1.65 μM. Aiming these applications, a biocatalysis approach
was used to obtain these HADs with optimal reactional conditions. In this study, reverse
micelles with immobilized Pseudomonas aeruginosa intact cells containing amidase
were used as a biocatalyst to catalyze the acyltransferase reaction of the corresponding
substrate amide and hydroxylamine to obtain various HAD and this was achieved for the
first time with yields of approximately 100%.
... This is because of unfavorable ionic conditions necessary for the formation of enzyme-substrate complex (Bhatia et al. 2013). Similar results were reported for amidases of Klebsiella pneumoniae NCTR 1 (Nawaz et al. 1996) and Bacillus smithii strain IITR6b2 (Agarwal et al. 2013) showing the maximal activity at neutral pH. ...
... The amidase from Alcaligenes sp. MTCC 10674 was reported to withstand 175 mM of product accumulated (Bhatia et al. 2013), Agarwal et al. (2013) used amidase from Bacillus smithii strain IITR6b2 for the synthesis of nicotinic acid hydroxamate, which has no inhibitory effect up to 536 mM product formed. Mehta et al. (2014) reported amidase of Geobacillus subterraneus RL-2a, wherein the enzyme has no inhibitory effect on the rate of product formation until 6 substrate feeds of 120 mM each against which 870 mM product was accumulated. ...
In this study, an amidohydrolase activity of amidase in whole cells of Rhodococcus sp. MTB5 has been used for the biotransformation of aromatic, monoheterocyclic and diheterocyclic amides to corresponding carboxylic acids. Benzoic acid, nicotinic acid and pyrazinoic acid are carboxylic acids which have wide industrial applications. The amidase of this strain is found to be inducible in nature. The biocatalytic conditions for amidase present in the whole cells of MTB5 were optimized against benzamide. The enzyme exhibited optimum activity in 50 mM potassium phosphate buffer pH 7.0. The optimum temperature and substrate concentrations for this enzyme were 50 °C and 50 mM, respectively. The enzyme was quite stable for more than 6 h at 30 °C. It showed substrate specificity against different amides, including aliphatic, aromatic and heterocyclic amides. Under optimized reaction conditions, the amidase is capable of converting 50 mM each of benzamide, nicotinamide and pyrazinamide to corresponding acids within 100, 160 and 120 min, respectively, using 5 mg dry cell mass (DCM) per mL of reaction mixture. The respective percent conversion of these amides was 95.02%, 98.00% and 98.44% achieved by whole cells. The amidase in whole cells can withstand as high as 383 mM concentration of product in a reaction mixture and above which it undergoes product feedback inhibition. The results of this study suggest that Rhodococcus sp. MTB5 amidase has the potential for large-scale production of carboxylic acids of industrial value.
... Besides their amide hydrolysis ability, most amidases were reported to exhibit acyl transfer activity and received great attention for synthesis of pharmaceutically important hydroxamic acids in the recent years [2,5,31]. The Abstract A novel amidase gene (bami) was cloned from Brevibacterium epidermidis ZJB-07021 by combination of degenerate PCR and high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR). ...
... It was also observed that the increased concentration of butyramide and hydroxylamine resulted in substrate inhibition, which was similar to amidase from Alcaligenes sp. MTCC 10674 [5] and Bacillus smithii strain IITR6b2 [2]. There were several undesirable reactions accompanying the biosynthesis of hydroxamic acids from amides, and by-products would be irreversibly produced by the initial amides hydrolysis and final hydroxamic acids hydrolysis [16]. ...
A novel amidase gene (bami) was cloned from Brevibacterium epidermidis ZJB-07021 by combination of degenerate PCR and high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR). The deduced amino acid sequence showed low identity (≤55 %) with other reported amidases. The bami gene was overexpressed in Escherichia coli, and the resultant inclusion bodies were refolded and purified to homogeneity with a recovery of 22.6 %. Bami exhibited a broad substrate spectrum towards aliphatic, aromatic and heterocyclic amides, and showed the highest acyl transfer activity towards butyramide with specific activity of 1331.0 ± 24.0 U mg−1. Kinetic analysis demonstrated that purified Bami exhibited high catalytic efficiency (414.9 mM−1 s−1) for acyl transfer of butyramide, with turnover number (K
cat) of 3569.0 s−1. Key parameters including pH, substrate/co-substrate concentration, reaction temperature and catalyst loading were investigated and the Bami showed maximum acyl transfer activity at 50 °C, pH 7.5. Enzymatic catalysis of 200 mM butyramide with 15 μg mL−1 purified Bami was completed in 15 min with a BHA yield of 88.1 % under optimized conditions. The results demonstrated the great potential of Bami for the production of a variety of hydroxamic acids.
... A number of different amide-degrading microorganisms have been earlier isolated from soil, but the isolate BR-1 exhibited a very high ATA for N-substituted aromatic amides, i.e. 100.64 ± 0.03 Umgdcm -1 , as compared with 9.9 Umg -1 , 24 Umg -1 and 16.1 Umgdcm -1 from the purified amidase of R. rhodochrous R312, Paracoccus sp. M1 and whole cells of Bacillus smithii strain IITR6b2 (Agarwal et al., 2013;Fournand et al., 1998;Shen et al., 2012). Pseudomonas putida BR-1 efficiently grew in a mineral salt medium (M5) resulting in considerable biomass yield, as well as the production of ATA. ...
Acyl transfer activity (ATA) of amidase transfers an acyl group of different amides to hydroxylamine to form the corresponding hydroxamic acid. Bacterial isolate BR-1 was isolated from cyanogenic plant Cirsium vulgare rhizosphere and identified as Pseudomonas putida BR-1 by 16S rDNA sequencing. This organism exhibited high ATA for the biotransformation of N-substituted aromatic amide to the corresponding hydroxamic acid. Optimization of media, tryptone (0.6%), inducer, pH 8.5, and a growth temperature 25 degrees C for 56 h, resulted in a 7-fold increase in ATA. Further, Response Surface Methodology (RSM) and multiple feeding approach (20 mM after 14 h) of inducer led to a 29% enhancement of ATA from this organism. The half life (t(1/2)) of this enzyme at 50 degrees C and 60 degrees C was 3 h and 1 h, respectively. The ATA of amidase of Pseudomonas putida BR-1 makes it a potential candidate for the production of a variety of N-substituted aromatic hydroxamic acid.
... A number of different amide-degrading microorganisms have been earlier isolated from soil, but the isolate BR-1 exhibited a very high ATA for N-substituted aromatic amides, i.e. 100.64 ± 0.03 Umgdcm -1 , as compared with 9.9 Umg -1 , 24 Umg -1 and 16.1 Umgdcm -1 from the purified amidase of R. rhodochrous R312, Paracoccus sp. M1 and whole cells of Bacillus smithii strain IITR6b2 (Agarwal et al., 2013;Fournand et al., 1998;Shen et al., 2012). Pseudomonas putida BR-1 efficiently grew in a mineral salt medium (M5) resulting in considerable biomass yield, as well as the production of ATA. ...
Amidase associated acyl transfer activity transfers
an acyl group of different amides to hydroxylamine
to form the corresponding hydroxamic acid.
Bacterial isolate BR-1, isolated from rhizosphere
of cyanogenic plant Cirsium vulgare and identified
as Pseudomonas putida BR-1 by 16S rDNA
sequencing, exhibited high acyl transfer activity for
N-substituted aromatic amide resulting in the formation
of the corresponding hydroxamic acid. Optimization
of media, nitrogen source tryptone
(0.6%), inducer isobutyronitrile, pH 8.5, growth
temperature 25∞C in 56 h resulted in a 7-fold increase
in acyl transfer activity. Further, Response
Surface Methodology (RSM) and multiple feeding
approach (20 mM after 14 h) of inducer led to a
29% enhancement of acyl transfer activity from
77.71 ± 0.02 Umgdcm–1 to 100.64 ± 0.03 Umgdcm–1
from this organism. The half life (t1/2) of this enzyme
at 50∞C and 60∞C was 3 h and 1 h, respectively.
The acyl transfer activity of amidase of Pseudomonas
putida BR-1 makes it a potential candidate
for the production of a variety of N-substituted
aromatic hydroxamic acid.
