Production and biochemical characterization of a type B ferulic acid esterase from Streptomyces ambofaciens

2009 ◽  
Vol 55 (6) ◽  
pp. 729-738 ◽  
Author(s):  
Fadi Kheder ◽  
Stéphane Delaunay ◽  
Ghassan Abo-Chameh ◽  
Cédric Paris ◽  
Lionel Muniglia ◽  
...  
Keyword(s):  
Ferulic Acid ◽  
Wheat Bran ◽  
Biochemical Characterization ◽  
Extracellular Enzyme ◽  
Type B ◽  
Ferulic Acid Esterase ◽  
Level Of Activity ◽  
Streptomyces Ambofaciens ◽  
Methyl Ferulate ◽  
First Time

For the first time, the presence of a ferulic acid esterase (FAE) was demonstrated in Streptomyces ambofaciens . This extracellular enzyme was produced on a range of lignocellulosic substrates. The maximal level of activity was detected in the presence of either destarched wheat bran or oat spelt xylan as the sole carbon source. We found that 1% (m/v) of destarched wheat bran was the optimal concentration to induce its production. With this inducer, no ferulic acid dimers were released from the cell wall by the produced FAE. Interestingly, rape cattle cake ( Brassica napus ), which does not contain esterified ferulic acid, was also shown to induce the production of the FAE from S. ambofaciens. The FAE was partially purified from the culture supernatant. The purified enzyme was optimally active at pH 7 and 40 °C. The substrate specificity of the FAE from S. ambofaciens was investigated: the highest activity was determined with methyl p-coumarate, methyl ferulate, and methyl cinnamate. Furthermore, the FAE required a certain distance between the benzene ring and the ester bond to be active. According to these biochemical characteristics, the FAE from S. ambofaciens has been classified as a type B FAE.

scholarly journals The Effect of Xylooligosaccharide, Xylan, and Whole Wheat Bran on the Human Gut Bacteria

2020 ◽  
Vol 11 ◽  
Author(s):  
Miao Chen ◽  
Shujun Liu ◽  
Khandaker Md. Sharif Uddin Imam ◽  
Lichao Sun ◽  
Yulu Wang ◽  
...  
Keyword(s):  
Ferulic Acid ◽  
Wheat Bran ◽  
Gut Bacteria ◽  
Venn Diagram ◽  
Metagenomic Sequencing ◽  
Dietary Fibers ◽  
Ferulic Acid Esterase ◽  
Human Gut ◽  
Whole Wheat

Wheat bran is a cereal rich in dietary fibers that have high levels of ferulic acid, which has prebiotic effects on the intestinal microbiota and the host. Herein we explored the effect of xylooligosaccharide, xylan, and whole wheat bran on the human gut bacteria and screened for potential ferulic acid esterase genes. Using in vitro fermentation, we analyzed the air pressure, pH-value, and short-chain fatty acid levels. We also performed 16S rRNA gene and metagenomic sequencing. A Venn diagram analysis revealed that 80% of the core operational taxonomic units (OTUs) were shared among the samples, and most of the xylooligosaccharide treatment core OTUs (319/333 OTUs) were shared with the other two treatments’ core OTUs. A significant difference analysis revealed that the relative abundance of Dorea, Bilophila, and Sulfurovum in wheat bran treatment was higher than that in xylan and xylooligosaccharide treatments. The clusters of orthologous groups of proteins functional composition of all samples was similar to the microbiota composition of the control. Using metagenomic sequencing, we revealed seven genes containing the conserved residues, Gly-X-Ser-X-Gly, and the catalytic triad, Ser-His-Asp, which are thus potential ferulic acid esterase genes. All the results indicate that xylan and/or xylooligosaccharide, the main dietary fibers in wheat bran, plays a major role in in vitro fermentation by the human gut microbiota.

scholarly journals Investigation of the Amycolatopsis sp. Strain ATCC 39116 Vanillin Dehydrogenase and Its Impact on the Biotechnical Production of Vanillin

2012 ◽  
Vol 79 (1) ◽  
pp. 81-90 ◽  
Author(s):  
Christian Fleige ◽  
Gunda Hansen ◽  
Jens Kroll ◽  
Alexander Steinbüchel
Keyword(s):  
Ferulic Acid ◽  
Vanillic Acid ◽  
Higher Plants ◽  
Strain Atcc ◽  
Biotechnological Production ◽  
Content Type ◽  
Level Of Activity ◽  
Bioinformatic Approaches ◽  
Using Data ◽  
First Time

ABSTRACTThe actinomyceteAmycolatopsissp. strain ATCC 39116 is capable of synthesizing large amounts of vanillin from ferulic acid, which is a natural cell wall component of higher plants. The desired intermediate vanillin is subject to undesired catabolism caused by the metabolic activity of a hitherto unknown vanillin dehydrogenase (VDHATCC 39116). In order to prevent the oxidation of vanillin to vanillic acid and thereby to obtain higher yields and concentrations of vanillin, the responsible vanillin dehydrogenase inAmycolatopsissp. ATCC 39116 was investigated for the first time by using data from our genome sequence analysis and further bioinformatic approaches. Thevdhgene was heterologously expressed inEscherichia coli, and the encoded vanillin dehydrogenase was characterized in detail. VDHATCC 39116was purified to apparent electrophoretic homogeneity and exhibited NAD+-dependent activity toward vanillin, coniferylaldehyde, cinnamaldehyde, and benzaldehyde. The enzyme showed its highest level of activity toward vanillin at pH 8.0 and at a temperature of 44°C. In a next step, a precisevdhdeletion mutant ofAmycolatopsissp. ATCC 39116 was generated. The mutant lost its ability to grow on vanillin and did not show vanillin dehydrogenase activity. A 2.3-times-higher vanillin concentration and a substantially reduced amount of vanillic acid occurred with theAmycolatopsissp. ATCC 39116 Δvdh::Kmrmutant when ferulic acid was provided for biotransformation in a cultivation experiment on a 2-liter-bioreactor scale. Based on these results and taking further metabolic engineering into account, theAmycolatopsissp. ATCC 39116 Δvdh::Kmrmutant represents an optimized and industrially applicable platform for the biotechnological production of natural vanillin.

scholarly journals A non-modular type B feruloyl esterase from Neurospora crassa exhibits concentration-dependent substrate inhibition

2003 ◽  
Vol 370 (2) ◽  
pp. 417-427 ◽  
Author(s):  
Valerie F. CREPIN ◽  
Craig B. FAULDS ◽  
Ian F. CONNERTON
Keyword(s):  
Cell Wall ◽  
Sugar Beet ◽  
Ferulic Acid ◽  
Neurospora Crassa ◽  
Plant Cell Wall ◽  
Substrate Inhibition ◽  
Type B ◽  
Ferulic Acid Esterase ◽  
Beet Pulp ◽  
Feruloyl Esterases

Feruloyl esterases, a subclass of the carboxylic acid esterases (EC 3.1.1.1), are able to hydrolyse the ester bond between the hydroxycinnamic acids and sugars present in the plant cell wall. The enzymes have been classified as type A or type B, based on their substrate specificity for aromatic moieties. We show that Neurospora crassa has the ability to produce multiple ferulic acid esterase activities depending upon the length of fermentation with either sugar beet pulp or wheat bran substrates. A gene identified on the basis of its expression on sugar beet pulp has been cloned and overexpressed in Pichia pastoris. The gene encodes a single-domain ferulic acid esterase, which represents the first report of a non-modular type B enzyme (fae-1 gene; GenBank accession no. AJ293029). The purified recombinant protein has been shown to exhibit concentration-dependent substrate inhibition (Km 0.048mM, Ki 2.5mM and Vmax 8.2units/mg against methyl 3,4-dihydroxycinnamate). The kinetic behaviour of the non-modular enzyme is discussed in terms of the diversity in the roles of the feruloyl esterases in the mobilization of plant cell wall materials and their respective modes of action.

scholarly journals Characterization of Feruloyl Esterase from Bacillus pumilus SK52.001 and Its Application in Ferulic Acid Production from De-Starched Wheat Bran

Foods ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1229
Author(s):  
Xiaoli Duan ◽  
Yiwei Dai ◽  
Tao Zhang
Keyword(s):  
Ferulic Acid ◽  
Wheat Bran ◽  
Bacillus Pumilus ◽  
Specific Activity ◽  
Added Value ◽  
Feruloyl Esterase ◽  
Waste Biomass ◽  
Biomass Degradation ◽  
Methyl Ferulate ◽  
Ferulic Acid Production

Feruloyl esterase (FAE; EC 3.1.1.73) catalyzes the hydrolysis of the 4-hydroxy-3-methoxycinnamoyl group in an esterified sugar to assist in waste biomass degradation or to release ferulic acid (FA). An FAE-producing strain was isolated from humus soil samples and identified as Bacillus pumilus SK52.001. The BpFAE gene from B. pumilus SK52.001 was speculated and heterogeneously expressed in Bacillus subtilis WB800 for the first time. The enzyme exists as a monomer with 303 amino acids and a molecular mass of 33.6 kDa. Its specific activity was 377.9 ± 10.3 U/ (mg protein), using methyl ferulate as a substrate. It displays an optimal alkaline pH of 9.0, an optimal temperature of 50 °C, and half-lives of 1434, 327, 235, and 68 min at 50, 55, 60, and 65 °C, respectively. Moreover, the purified BpFAE released 4.98% FA of the alkali-acidic extractable FA from de-starched wheat bran (DSWB). When the DSWB was enzymatically degraded by the synergistic effect of the BpFAE and commercial xylanase, the FA amount reached 49.47%. It suggested that the alkaline BpFAE from B. pumilus SK52.001, which was heterologously expressed in B. subtilis WB800, possesses great potential for biomass degradation and achieving high-added value FA production from food by-products.

scholarly journals Molecular characterization of cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene and frequency of blood types in stray cats of İzmir, Turkey

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Hüseyin Can ◽  
Sedef Erkunt Alak ◽  
Ahmet Efe Köseoğlu ◽  
Umut Şahar ◽  
Berna Bostanbaş ◽  
...  
Keyword(s):  
Type A ◽  
Type B ◽  
Exon 2 ◽  
Blood Types ◽  
Acetylneuraminic Acid ◽  
Stray Cats ◽  
High Prevalence ◽  
Heterozygous Form ◽  
Type Ab ◽  
First Time

Abstract Background Cytidine monophospho-n-acetylneuraminic acid hydroxylase (CMAH) gene associated with blood groups in cats encodes CMAH enzyme that converts Neu5Ac to Neu5Gc. Although variations in CMAH gene of pedigree cats have been revealed, the presence/lack of them in non-pedigree stray cats is unknown. Therefore, the present study aimed to investigate the variations in CMAH gene and the quantity of Neu5Ac and Neu5Gc on erythrocytes of non-pedigree stray cats (n:12) living in İzmir, Turkey. Also, the frequency of blood types was determined in 76 stray cats including 12 cats that were used for CMAH and Neu5A/Neu5Gc analysis. Results In total, 14 SNPs were detected in 5’UTR as well as in exon 2, 4, 9, 10, 11 and 12 of CMAH gene. Among these SNPs, -495 C > T in 5’UTR was detected for the first time as heterozygous in type A and AB cats, and homozygous and heterozygous in type B cats. The remaining 13 that have been detected in previous studies were also found as homozygous or heterozygous. Both Neu5Gc and Neu5Ac were detected in type A and AB cats. In type B cats, only Neu5Ac was detected. Among two type AB cats, the level of Neu5Ac was found higher in cat carrying heterozygous form (T/C) of 1392T > C. The prevalence of type B cats (67.1 %) was higher than others. Conclusions The presence of a new SNP as well as previous SNPs indicates that more variations can be found in stray cats with a more comprehensive study in the future. Also, the high prevalence of type B cats demonstrates the possible risk of neonatal isoerythrolysis among stray cats living in İzmir, Turkey.

Microarray analysis of the transcriptome of the subfornical organ in the rat: regulation by fluid and food deprivation

2008 ◽  
Vol 295 (6) ◽  
pp. R1914-R1920 ◽  
Author(s):  
Charles Hindmarch ◽  
Mark Fry ◽  
Song T. Yao ◽  
Pauline M. Smith ◽  
David Murphy ◽  
...  
Keyword(s):  
Gene Expression ◽  
Statistical Analysis ◽  
Food Deprivation ◽  
Subfornical Organ ◽  
Microarray Technology ◽  
Type B ◽  
Bdnf Expression ◽  
Experimental Conditions ◽  
Calcium Sensing ◽  
First Time

We have employed microarray technology using Affymetrix 230 2.0 genome chips to initially catalog the transcriptome of the subfornical organ (SFO) under control conditions and to also evaluate the changes (common and differential) in gene expression induced by the challenges of fluid and food deprivation. We have identified a total of 17,293 genes tagged as present in one of our three experimental conditions, transcripts, which were then used as the basis for further filtering and statistical analysis. In total, the expression of 46 genes was changed in the SFO following dehydration compared with control animals (22 upregulated and 24 downregulated), with the largest change being the greater than fivefold increase in brain-derived neurotrophic factor (BDNF) expression, while significant changes in the expression of the calcium-sensing (upregulated) and apelin (downregulated) receptors were also reported. In contrast, food deprivation caused greater than twofold changes in a total of 687 transcripts (222 upregulated and 465 downregulated), including significant reductions in vasopressin, oxytocin, promelanin concentrating hormone, cocaine amphetamine-related transcript (CART), and the endothelin type B receptor, as well as increases in the expression of the GABAB receptor. Of these regulated transcripts, we identified 37 that are commonly regulated by fasting and dehydration, nine that were uniquely regulated by dehydration, and 650 that are uniquely regulated by fasting. We also found five transcripts that were differentially regulated by fasting and dehydration including BDNF and CART. In these studies we have for the first time described the transcriptome of the rat SFO and have in addition identified genes, the expression of which is significantly modified by either water or food deprivation.

scholarly journals Biochemical Characterization of Individual Components of the Allochromatium vinosum DsrMKJOP Transmembrane Complex Aids Understanding of Complex Function In Vivo

2010 ◽  
Vol 192 (24) ◽  
pp. 6369-6377 ◽  
Author(s):  
Fabian Grein ◽  
Inês A. C. Pereira ◽  
Christiane Dahl
Keyword(s):  
Biochemical Characterization ◽  
Sulfur Metabolism ◽  
Complex Function ◽  
Redox Potentials ◽  
Allochromatium Vinosum ◽  
Purple Sulfur Bacterium ◽  
Membrane Anchoring ◽  
Integral Proteins ◽  
First Time

ABSTRACT The DsrMKJOP transmembrane complex has a most important function in dissimilatory sulfur metabolism and consists of cytoplasmic, periplasmic, and membrane integral proteins carrying FeS centers and b- and c-type cytochromes as cofactors. In this study, the complex was isolated from the purple sulfur bacterium Allochromatium vinosum and individual components were characterized as recombinant proteins. The two integral membrane proteins DsrM and DsrP were successfully produced in Escherichia coli C43(DE3) and C41(DE3), respectively. DsrM was identified as a diheme cytochrome b, and the two hemes were found to be in low-spin state. Their midpoint redox potentials were determined to be +60 and +110 mV. Although no hemes were predicted for DsrP, it was also clearly identified as a b-type cytochrome. To the best of our knowledge, this is the first time that heme binding has been experimentally proven for a member of the NrfD protein family. Both cytochromes were partly reduced after addition of a menaquinol analogue, suggesting interaction with quinones in vivo. DsrO and DsrK were both experimentally proven to be FeS-containing proteins. In addition, DsrK was shown to be membrane associated, and we propose a monotopic membrane anchoring for this protein. Coelution assays provide support for the proposed interaction of DsrK with the soluble cytoplasmic protein DsrC, which might be its substrate. A model for the function of DsrMKJOP in the purple sulfur bacterium A. vinosum is presented.

scholarly journals Study on a Fermented Whole Wheat: Phenolic Content, Activity on PTP1B Enzyme and In Vitro Prebiotic Properties

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1120 ◽  
Author(s):  
Diletta Balli ◽  
Maria Bellumori ◽  
Paolo Paoli ◽  
Giuseppe Pieraccini ◽  
Monica Di Paola ◽  
...  
Keyword(s):  
Lactobacillus Reuteri ◽  
Tyrosine Phosphatase ◽  
Bacterial Species ◽  
Food Ingredient ◽  
Microbial Composition ◽  
Acidic Hydrolysis ◽  
Inhibitory Power ◽  
Methyl Ferulate ◽  
First Time

Fermented cereals, staple foods in Asia and Africa, are recently receiving a growing interest in Western countries. The object of this work is the characterization of a fermented wheat used as a food ingredient and dietary supplement. To this aim, the phenolic composition, the activity on protein tyrosine phosphatase 1B (PTP1B), an enzyme overexpressed in type-II diabetes, the in vitro prebiotic properties on Lactobacillus reuteri and the microbial composition were investigated. Basic and acidic hydrolysis were tested for an exhaustive recovery of bound phenols: the acidic hydrolysis gave best yields. Methyl ferulate and neocarlinoside were identified for the first time in wheat. The inhibitory power of the extracts of several batches were investigated on PTP1B enzyme. The product was not able to inhibit the enzyme, otherwise, for the first time, a complete inhibition was observed for schaftoside, a major C-flavonoid of wheat. The microbial composition was assessed identifying Lactobacillus, Enterococcus, and Pediococcus as the main bacterial species. The fermented wheat was a suitable substrate for the grown of L. reuteri, recognized for its health properties in the human gut. The proposed method for phenols is easier compared to those based on strong basic hydrolysis; our results assessed the bound phenols as the major fraction, differently from that suggested by the literature for fermented cereals.

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