scholarly journals The commonness and difference among the Lactobacillus feruloyl esterases expressed in Escherichia coli

2020 ◽  
Author(s):  
Zhenshang Xu ◽  
Jian Kong ◽  
Susu Zhang ◽  
Ting Wang ◽  
Xinli Liu
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Wheat Bran ◽  
Agricultural Waste ◽  
Feruloyl Esterase ◽  
E Coli ◽  
One Step ◽  
Feruloyl Esterases ◽  
Extracellular Environment ◽  
Extracellular Activity

Abstract Background: Construction of recombinant Escherichia coli strains carrying feruloyl esterase genes for secretory expression offers an attractive way to facilitate enzyme purification and one-step produce ferulic acid from agricultural waste. It was found that the Lactobacillus crispatus feruloyl esterase could be secreted into extracellular environment of E. coli. Whether other Lactobacillus feruloyl esterases share the same secretory characteristic is worth investigation.Results: A total of ten feruloyl esterases derived from nine Lactobacillus species were used to analyze their commonness and compare their difference when heterologously expressed in E. coli BL21 (DE3). Extracellular activity determination showed all these Lactobacillus feruloyl esterases could be secreted out of E. coli cells. However, protein analysis indicated that they could be classified as three types. The first type presented a low secretion level, including feruloyl esterases derived from Lb. acidophilus and Lb. johnsonii. The second type showed a high secretion level, including feruloyl esterases derived from Lb. amylovorus, Lb. crispatus, Lb. gasseri and Lb. helveticus. The third type also behaved a high secretion level but easy degradation, including feruloyl esterases derived from Lb. farciminis, Lb. fermentum and Lb. reuteri. Moreover, these recombinant E. coli strains could directly release ferulic acid from de-starched wheat bran. Conclusions: Recombinant E. coli strains expressing feruloyl esterase of Lb. amylovorus, Lb. crispatus and Lb. helveticus displayed high secretion level and stable extracellular activity. Furthermore, the highest yield of ferulic acid was 140 µg on the basis of 0.1 g de-starched wheat bran after 72 h cultivation of E. coli expressing Lb. amylovorus feruloyl esterase. These results provided a solid basis for the production of feruloyl esterase and ferulic acid.

scholarly journals Comparison of Enzyme Secretion and Ferulic Acid Production by Escherichia coli Expressing Different Lactobacillus Feruloyl Esterases

2020 ◽  
Vol 11 ◽  
Author(s):  
Zhenshang Xu ◽  
Jian Kong ◽  
Susu Zhang ◽  
Ting Wang ◽  
Xinli Liu
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Acid Production ◽  
Lactobacillus Reuteri ◽  
Agricultural Waste ◽  
Lactobacillus Helveticus ◽  
Feruloyl Esterase ◽  
E Coli ◽  
Feruloyl Esterases ◽  
Ferulic Acid Production

Construction of recombinant Escherichia coli strains carrying feruloyl esterase genes for secretory expression offers an attractive way to facilitate enzyme purification and one-step production of ferulic acid from agricultural waste. A total of 10 feruloyl esterases derived from nine Lactobacillus species were expressed in E. coli BL21 (DE3) to investigate their secretion and ferulic acid production. Extracellular activity determination showed all these Lactobacillus feruloyl esterases could be secreted out of E. coli cells. However, protein analysis indicated that they could be classified as three types. The first type presented a low secretion level, including feruloyl esterases derived from Lactobacillus acidophilus and Lactobacillus johnsonii. The second type showed a high secretion level, including feruloyl esterases derived from Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus gasseri, and Lactobacillus helveticus. The third type also behaved a high secretion level but easy degradation, including feruloyl esterases derived from Lactobacillus farciminis, Lactobacillus fermentum, and Lactobacillus reuteri. Moreover, these recombinant E. coli strains could directly release ferulic acid from agricultural waste. The highest yield was 140 μg on the basis of 0.1 g de-starched wheat bran by using E. coli expressed L. amylovorus feruloyl esterase. These results provided a solid basis for the production of feruloyl esterase and ferulic acid.

scholarly journals One-step fermentation for producing xylo-oligosaccharides from wheat bran by recombinant Escherichia coli containing an alkaline xylanase

2021 ◽  
Author(s):  
Jiawen Liu ◽  
Cong Liu ◽  
Shilei Qiao ◽  
Zhen Dong ◽  
Di Sun ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Wheat Bran ◽  
Xylanase Activity ◽  
Maximal Activity ◽  
Fermentation Medium ◽  
Growth Conditions ◽  
Glycoside Hydrolase Family ◽  
E Coli ◽  
Response Surface Optimization ◽  
One Step

Abstract One-step fermentation is a cheap way to produce xylo-oligosaccharides (XOS), where production of xylanases and XOS is integrated into a single process. In spite of having cost advantage, one-step fermentation is still short in yield so far due to the limited exploration. To cope with this issue, production of XOS from wheat bran by recombinant Escherichia coli through one-step fermentation was investigated here. A xylanase gene belonging to glycoside hydrolase family 11 of Bacillus agaradhaerens was employed to construct recombinant E. coli. This xylanase showed maximal activity at 60°C and pH 8.0. Its activity retained more than 60% after incubation at 70°C for 4 hours, showing a good stability. The recombinant E. coli successfully secreted xylanases that directly hydrolyzed wheat bran to XOS in fermentation medium. The generated XOS consisted of xylose, xylobiose and xylotriose accounting for 23.1%, 37.3% and 39.6%, respectively. Wheat bran concentration was found to be the most crucial factor affecting XOS production. The yield reached 5.3 mg/mL at 10% of wheat bran, which is higher than previous reports employing one-step fermentation. Nitrogen source type could also affect XOS yield by changing extracellular xylanase activity, and glycine was found to be the best one for fermentation. Optimal fermentation conditions were finally studied by response surface optimization. The maximal yield emerged at 44.3°C, pH 7.98, which is affected by characteristics of the xylanase and growth conditions of E. coli. This work indicates that the integrated fermentation using recombinant E. coli is highly competitive in cost and yield for production of XOS.

scholarly journals Novel Feruloyl Esterase from Lactobacillus fermentum NRRL B-1932 and Analysis of the Recombinant Enzyme Produced in Escherichia coli

2016 ◽  
Vol 82 (17) ◽  
pp. 5068-5076 ◽  
Author(s):  
Siqing Liu ◽  
Kenneth M. Bischoff ◽  
Amber M. Anderson ◽  
Joseph O. Rich
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Animal Feed ◽  
Lactobacillus Fermentum ◽  
Biofuel Production ◽  
Artificial Substrates ◽  
Feruloyl Esterase ◽  
Content Type ◽  
Ethyl Ferulate ◽  
E Coli

ABSTRACTA total of 33Lactobacillusstrains were screened for feruloyl esterase (FE) activity using agar plates containing ethyl ferulate as the sole carbon source, andLactobacillus fermentumNRRL B-1932 demonstrated the strongest FE activity among a dozen species showing a clearing zone on the opaque plate containing ethyl ferulate. FE activities were monitored using high-performance liquid chromatography with an acetonitrile-trifluoroacetic acid gradient. To produce sufficient purified FE fromL. fermentumstrain NRRL B-1932 (LfFE), the cDNA encoding LfFE (Lffae) was amplified and cloned by using available closely related genome sequences and overexpressed inEscherichia coli. A 29.6-kDa LfFE protein was detected from the protein extract ofE. coliBL21(pLysS) carrying pET28bLffaeupon IPTG (isopropyl-β-d-thiogalactopyranoside) induction. The recombinant LfFE containing a polyhistidine tag was purified by nickel-nitrilotriacetic acid affinity resin. The purified LfFE showed strong activities against several artificial substrates, includingp-nitrophenyl acetate and 4-methylumbelliferylp-trimethylammoniocinnamate chloride. The optimum pH and temperature of the recombinant LfFE were around 6.5 and 37°C, respectively, as determined using either crude or purified recombinant LfFE. This study will be essential for the production of the LfFE inE. colion a larger scale that could not be readily achieved byL. fermentumfermentation.IMPORTANCEThe production of feruloyl esterase (FE) fromLactobacillus fermentumNRRL B-1932 reported in this study will have immense potential commercial applications not only in biofuel production but also in pharmaceutical, polymer, oleo chemical, cosmetic additive, and detergent industries, as well as human health-related applications, including food flavoring, functional foods, probiotic agents, preventive medicine, and animal feed. Given the essential role FE plays in the production of hydroxycinnamic acids and ferulic acid, plus the generally regarded as safe status of lactobacilli, which therefore have less regulatory concerns, LfFE from the probioticL. fermentumreported in this work can be directly used for increased production of high-value hydroxycinnamates and ferulic acid from natural or synthetic carbon sources.

One-Step Preparation of Competent E. coli: Transformation and Storage of Bacterial Cells in the Same Solution

2021 ◽  
Vol 2021 (11) ◽  
pp. pdb.prot101212 ◽  
Author(s):  
Michael R. Green ◽  
Joseph Sambrook
Keyword(s):  
Escherichia Coli ◽  
Convenient Method ◽  
Plasmid Dna ◽  
Transformation Efficiency ◽  
Bacterial Cells ◽  
E Coli ◽  
One Step ◽  
And Storage

This protocol describes a convenient method for the preparation, use, and storage of competent Escherichia coli. The reported transformation efficiency of this method is ∼5 × 107 transformants/µg of plasmid DNA.

scholarly journals PSV-41 Antibacterial activity of ferulic acid and sodium chlorate against Escherichia coli F18 and K88 during incubations with porcine fecal bacteria

2019 ◽  
Vol 97 (Supplement_3) ◽  
pp. 296-297 ◽  
Author(s):  
Claudio Arzola ◽  
Elizabeth Latham ◽  
Robin Anderson ◽  
Jaime Salinas-Chavira ◽  
Yamicela Castillo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Sodium Chlorate ◽  
Fecal Bacteria ◽  
Aerobic Incubation ◽  
E Coli ◽  
Mueller Hinton ◽  
Fecal Suspension ◽  
Porcine Feces ◽  
Mueller Hinton Broth

Abstract The influence of ferulic acid (FA) and sodium chlorate (SC) was evaluated in two trials on the growth of Escherichia coli F18 and K88 (F18 and K88) incubated with porcine fecal bacteria. Treatments were 2 levels of FA (0 and 5 mg/mL) and 2 levels of SC (0 and 10 mM/mL). In trial one, ½-strength Mueller Hinton broth mixed with porcine feces (0.5% w/v) was inoculated with a novobiocin and naladixic acid resistant F18-strain. This fecal suspension was transferred to tubes (3/treatment) and anaerobically incubated at 39 oC for enumeration at 0, 6 and 24 h using MacConkey agar supplemented with novobiocin and naladixic acid with aerobic incubation at 37 oC. An interaction (FA x SC) at 6 and 24 h was observed (P < 0.01). At 6 h of incubation, SC alone or combined with FA had the lowest counts (P < 0.05); FA alone was lower than control but higher than SC or SC+FA (P < 0.05). At 24 h, FA alone or combined with SC had the lowest counts (P < 0.05); SC was lower than control but higher than FA or SC+FA (P < 0.05). In trial 2 were used the same procedures of trial 1, except that K88 was used. There was an interaction at 6 h (P < 0.01) where the lowest counts were in FA+SC (P < 0.05). SC alone or FA alone were lower than control but higher than SC+FA (P < 0.05). There was no interaction at 24 h (P = 0.16), where FA reduced the K88 counts (P < 0.01), however it was not affected by SC (P = 0.12). In conclusion, SC reduced E. coli counts; however, at 24 h of incubation greater reductions were observed when FA alone or combined with SC was added into the incubation fluid with porcine feces.

scholarly journals PURIFICATION OF DUG WELL WATER FROM Escherichia coli BY USING CARBON OF RICE HUSK

2021 ◽  
Vol 8 (2) ◽  
pp. 96
Author(s):  
Hagar Karlani ◽  
Fahrul Fahrul ◽  
Maria M Meiwati ◽  
Herianus Manimoy ◽  
Loth Botahala
Keyword(s):  
Escherichia Coli ◽  
Rice Husk ◽  
Water Purification ◽  
Agricultural Waste ◽  
Well Water ◽  
Clean Water ◽  
E Coli ◽  
Dug Wells ◽  
Dug Well

Clean water is one of the human's main needs. Dug well water in Tombang, used for various needs. However, the dug well water has been polluted by E. Coli bacteria which is suspected to be due to the disposal of rubbish and waste carelessly and also the distance of latrines and dug wells that are very close together. Therefore it is important to do water purification using agricultural waste that is not utilized. The analysis showed that the purification of dug well water in Tombang with rice husk charcoal had reduced E. Coli bacteria by 55.6% with MPN pattern 1-0-3 so that the MPN value was 11/100 mL of sample.

scholarly journals Construction of an Artificial Biosynthetic Pathway for the Styrylpyrone Compound 11-Methoxy-Bisnoryangonin Produced in Engineered Escherichia coli

2021 ◽  
Vol 12 ◽  
Author(s):  
Kyung Taek Heo ◽  
Byeongsan Lee ◽  
Jae-Hyuk Jang ◽  
Jung-Oh Ahn ◽  
Young-Soo Hong
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Biosynthetic Pathway ◽  
De Novo ◽  
Chain Elongation ◽  
Piper Nigrum ◽  
Glucose Medium ◽  
E Coli ◽  
Acid Catalyzed ◽  
Simple Sugar

A cDNA clone (named pnpks), which shows high homology to the known chalcone synthase (CHS)-like type III PKS, was obtained from the leaves of Piper nigrum. The PnPKS protein with ferulic acid catalyzed lactonization instead of chalcone or stilbene formation. The new product was characterized as a styrylpyrone, 11-methoxy-bisnoryangonin, which is the lactonization compound of a linear triketide formed as the reaction product of PnPKS protein with ferulic acid. These results show that pnpks encodes a styrylpyrone synthase (SPS)-like PKS that catalyzes two-chain elongation with feruloyl CoA-linked starter substrates. Although these styrylpyrone compounds are promising for use in human healthcare, they are mainly obtained by extraction from raw plant or mushroom sources. For de novo synthesis of 11-methoxy-bisnoryangonin in the heterologous host Escherichia coli from a simple sugar as a starter, the artificial biosynthetic pathway contained five genes: optal, sam5, com, and 4cl2nt, along with the pnpks gene. The engineered L-tyrosine overproducing E. coli ∆COS1 strain, in which five biosynthetic genes were cloned into two vectors, pET-opT5M and pET22-4P, was cultured for 24 h in a minimal glucose medium containing ampicillin and kanamycin. As a result, 11-methoxy-bisnoryangonin production of up to 52.8 mg/L was achieved, which is approximately 8.5-fold higher than that in the parental E. coli strain harboring a plasmid for 11-methoxy-bisnoryangonin biosynthesis. As a potential styrylpyrone compound, 11-methoxy-bisnoryangonin, was successfully produced in E. coli from a simple glucose medium, and its production titer was also increased using engineered strains. This study provides a useful reference for establishing the biological manufacture of styrylpyrone compounds.

scholarly journals Ferulic Acid production by metabolically engineered Escherichia coli

2021 ◽  
Author(s):  
Huajun Lv ◽  
Ying Zhang ◽  
Jie Shao ◽  
Haili Liu ◽  
Yong Wang
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Caffeic Acid ◽  
Pyridine Nucleotide ◽  
Promoter Strength ◽  
T7 Promoter ◽  
Nadph Regeneration ◽  
E Coli ◽  
Ferulic Acid Production ◽  
K 12

Abstract Ferulic acid (p-hydroxy-3-methoxycinnamic acid, FA) is a natural active substance present in plant cell walls, with antioxidant, anticancer, antithrombotic and other properties; it is widely used in medicine, food, and cosmetics areas. Production of FA by eco-friendly bioprocess is of great potential. In this study, FA was biosynthesized by metabolically engineered Escherichia coli. As the first step, the genes tal (encoding Tyrosine ammonia-lyase, RsTAL) from Rhodobacter sphaeroides, sam5 (encoding p - coumarate 3-hydroxylase, SeSAM5) from Saccharothrix espanaensis and comt (encoding Caffeic acid O-methytransferase, TaCM) from Triticum aestivum were cloned in an operon on the pET plasmid backbone, E. coli strain containing this construction was proved to produce FA from L-tyrosine successfully, and confirmed the function of TaCM as Caffeic acid O-methytransferase. Fermentation results revealed JM109(DE3) as more suitable host cell for FA production than BL21(DE3). After that the genes expression strength of FA pathway were optimized by tuning of promoter strength (T7 promoter or T5 promoter) and copy number (pBR322 ori or p15a ori), and the combination p15a-T5 works best. To further improve FA production, E.coli native pntAB, encoding pyridine nucleotide transhydrogenase, was selected from five NADPH regeneration genes to supplement redox cofactor NADPH for converting p-coumaric acid into caffeic acid in FA biosynthesis process. Sequentially, to further convert caffeic acid into FA, a non-native methionine kinase (MetK from Streptomyces spectabilis) was also over expressed. Based on the flask fermentation data which shows that the engineered E. coli strain produced 212 mg/L of FA with 11.8 mg/L caffeic acid residue, it could be concluded that it is the highest yield of FA achieved by E.coli K-12 strains reported to the best of our knowledge.

scholarly journals Influence of sodium chlorate, ferulic acid, and essential oils on Escherichia coli and porcine fecal microbiota

2020 ◽  
Vol 98 (3) ◽  
Author(s):  
Claudio Arzola-Alvarez ◽  
Michael E Hume ◽  
Robin C Anderson ◽  
Elizabeth A Latham ◽  
Oscar Ruiz-Barrera ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ferulic Acid ◽  
Essential Oils ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Gene Sequencing ◽  
Sodium Chlorate ◽  
The Other ◽  
Enterotoxigenic Escherichia Coli ◽  
Similarity Coefficients ◽  
E Coli

Abstract The influence of sodium chlorate (SC), ferulic acid (FA), and essential oils (EO) was examined on the survivability of two porcine diarrhetic enterotoxigenic Escherichia coli (ETEC) strains (F18 and K88) and populations of porcine fecal bacteria. Fecal bacterial populations were examined by denaturing gradient gel electrophoresis (DGGE) and identification by 16S gene sequencing. The treatments were control (no additives), 10 mM SC, 2.5 mg FA /mL, a 1.5% vol/vol solution of an EO mixture as well as mixtures of EO + SC, EO + FA, and FA + SC at each of the aforementioned concentrations. EO were a commercial blend of oregano oil and cinnamon oil with water and citric acid. Freshly collected porcine feces in half-strength Mueller Hinton broth was inoculated with E. coli F18 (Trial 1) or E. coli K88 (Trial 2). The fecal-E. coli suspensions were transferred to crimp top tubes preloaded with the treatment compounds. Quantitative enumeration was at 0, 6, and 24 h. All treatments reduced (P < 0.05) the counts of E. coli F18 at 6 and 24 h. With the exception of similarity coefficient (%SC), all the other treatments reduced (P < 0.05) the K88 counts at 24 h. The most effective treatments to reduce the F18 and K88 CFU numbers were those containing EO. Results of DGGE revealed that Dice percentage similarity coefficients (%SC) of bacterial profiles among treatment groups varied from 81.3% to 100%SC. The results of gene sequencing showed that, except for SC at 24 h, all the other treatments reduced the counts of the family Enterobacteriaceae, while Lactobacillaceae and Ruminococcaceae increased and Clostridiaceae decreased in all treatments. In conclusion, all treatments were effective in reducing the ETEC, but EO mixture was the most effective. The porcine microbial communities may be influenced by the studied treatments.

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