scholarly journals High-Frequency Occurrence of Surfactin Monomethyl Isoforms in the Ferment Broth of a Bacillus subtilis Strain Revealed by Ion Trap Mass Spectrometry

Molecules ◽  
2018 ◽  
Vol 23 (9) ◽  
pp. 2224 ◽  
Author(s):  
Anita Kecskeméti ◽  
Attila Bartal ◽  
Bettina Bóka ◽  
László Kredics ◽  
László Manczinger ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Amino Acid ◽  
Aspartic Acid ◽  
Ion Trap ◽  
Biological Activities ◽  
Biological Effects ◽  
Amino Acid Position ◽  
Molecular Ions ◽  
Bacillus Species ◽  
Subtilis Strain

Surfactins are cyclic lipopeptides consisting of a β-hydroxy fatty acid of various chain length and a peptide ring of seven amino acids linked together by a lactone bridge, forming the cyclic structure of the peptide chain. These compounds are produced mainly by Bacillus species and possess numerous biological effects such as antimicrobial (antiviral, antibacterial, and antifungal) activities. A mixture of surfactins extracted from Bacillus subtilis strain SZMC 6179J was examined by HPLC-ESI-IT-MS technique, enhancing their separation to reveal novel lipopeptide varieties with higher masses and to characterize their structures. During the MS2 spectra analyses of their sodiated molecular ions [M + Na]+, a previously rarely encountered group of surfactins was detected and two novel types of the group were discovered containing methyl esterified aspartic acid residue in their fifth amino acid position. The relative amounts of these monomethyl isoforms exceeded 35% of the produced surfactin in total. In the m/z value of 1114, all the detected isoforms possessed aspartic acid 4-methyl ester residue in their fifth amino acid position (C17-[Lxx4, AME5], C18-[AME5]), offering an opportunity to separate a pure fraction of the compound and to study its biological activities in the future.

scholarly journals Effects of Different Cultivation Parameters on the Production of Surfactin Variants by a Bacillus subtilis Strain

Molecules ◽  
2018 ◽  
Vol 23 (10) ◽  
pp. 2675 ◽  
Author(s):  
Attila Bartal ◽  
Aruna Vigneshwari ◽  
Bettina Bóka ◽  
Mónika Vörös ◽  
István Takács ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Metal Ions ◽  
Biological Activities ◽  
Wide Spectrum ◽  
Carbon Sources ◽  
Bacillus Species ◽  
Subtilis Strain ◽  
Fatty Acid Chain ◽  
Characteristic Features ◽  
Surfactin Production

Surfactins are lipopeptide-type biosurfactants produced mainly by Bacillus species, consisting of a peptide loop of seven amino acids and a hydrophobic fatty acid chain (C12–C16). These molecules have been proven to exhibit various biological activities; thus, their therapeutic and environmental applications are considered. Within the surfactin lipopeptide family, there is a wide spectrum of different homologues and isomers; to date, more than 30 variants have been described. Since the newest members of these lipopeptides were described recently, there is no information that is available on their characteristic features, e.g., the dependence of their production from different cultivation parameters. This study examined the effects of both the different carbon sources and various metal ions on the surfactin production of a selected B. subtilis strain. Among the applied carbon sources, fructose and xylose had the highest impacts on the ratio of the different variants, regarding both the peptide sequences and the lengths of the fatty acids. Furthermore, the application of metal ions Mn2+, Cu2+ and Ni2+ in the media completely changed the surfactin variant compositions of the fermenting broths leading to the appearance of methyl esterified surfactin forms, and resulted in the appearance of novel surfactin variants with fatty acid chains containing no more than 11 carbon atoms.

scholarly journals Sequence of the N-terminal half of Bacillus amyloliquefaciens α-amylase

1980 ◽  
Vol 185 (2) ◽  
pp. 387-395 ◽  
Author(s):  
H Chung ◽  
F Friedberg
Keyword(s):  
Amino Acids ◽  
Bacillus Subtilis ◽  
Amino Acid ◽  
Aspartic Acid ◽  
Primary Structure ◽  
Bacillus Amyloliquefaciens ◽  
Proteolytic Enzymes ◽  
Alpha Amylase ◽  
Sequence Determination ◽  
N Terminus

Bacillus amyloliquefaciens alpha-amylase (1,4-alpha-D-glucan glucanohydrolase. EC 3.2.1.1), which is commercially supplied as ‘Bacillus subtilis alpha-amylase’ does not cross-react immunologically with B. subtilis alpha-amylase. This enzyme (from B. amyloliquefaciens) was cleaved by treatment with CNBr into seven fragments. Peptide A was selected for sequence determination. It is the longest one, containing 185 amino acids (i.e. approx. 50% of the total molecule) and connects to the hexapeptide of the N-terminus. Its primary structure was aligned by use of various proteolytic enzymes. The sequence of amino acids 181-184 is identical with that of amino acids 14-17 of the alpha-amylase isolated from B. subtilis (except that amino acid 183 is asparagine rather than aspartic acid).

Optimization and Partial Characterization of Biosurfactants Produced by Bacillus Species and Their Potential for Ex-Situ Enhanced Oil Recovery

SPE Journal ◽  
2011 ◽  
Vol 16 (03) ◽  
pp. 672-682 ◽  
Author(s):  
H.. Al-Sulaimani ◽  
Y.. Al-Wahaibi ◽  
S.. Al-Bahry ◽  
A.. Elshafie ◽  
A.. Al-Bemani ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Enhanced Oil Recovery ◽  
Oil Recovery ◽  
Contaminated Soils ◽  
Carbon Sources ◽  
Ex Situ ◽  
Bacillus Species ◽  
Subtilis Strain ◽  
Ft Ir ◽  
Ph Range

Summary This study aims to test the potential of microbial enhanced oil recovery (MEOR) as an effective alternative in Omani oil fields. In this study, biosurfactants produced by Bacillus licheniformis and Bacillus subtilis strains isolated from oil-contaminated soils from different parts of the Sultanate of Oman were investigated. Eight different minimal production media using different sugars as carbon sources were tested on three of the strains to select the medium that maximized the production of biosurfactants which were indicated by the interfacial tension (IFT) reduction. All isolates were tested on their potential media to screen for the best biosurfactant producer among the available strains. It was found that Bacillus subtilis Strain W19 gave the maximum IFT reduction (46.6 mN/m to 3.28 mN/m) in 16 hours of incubation when grown in a minimal medium containing glucose. The yield of the biosurfactant produced by B. subtilis W19 was 2.5g/L. Critical micelle concentration (CMC) was measured to be 0.4 g/L. The biosurfactant was partially characterized by FT-IR analysis, in which the peaks obtained imply the presence of aliphatic hydrocarbons as well as a peptide-like moiety in the biosurfactant. The cell-free biosurfactant broth produced from Strain W19 was found to be stable over a pH range from 6 to 10 and was most effective at 7. It also showed no loss in surface activity when subjected to various temperatures (60, 80, and 120°C). The biosurfactant also retained almost 60% of its activity even in a high-saline environment up to 20% NaCl (w/v). Further studies were conducted to test the interaction of biosurfactant produced by B. subtilis W19 with porous media in coreflooding experiments as a tertiary-recovery stage. The results showed high potential of using this bacterium during ex-situ MEOR applications in which a total of 23% of residual oil was produced after biosurfactant and concentrated-biosurfactant injection.

Cloning, Expression and Characterization of a Lipase from Bacillus Subtilis Strain I4 with Potential Application in Biodiesel Production

2013 ◽  
Vol 291-294 ◽  
pp. 243-248 ◽  
Author(s):  
Zhen Rong Xie ◽  
Xue Lin Zhang ◽  
Jun Mei Ding ◽  
Jun Jun Li ◽  
Yun Juan Yang ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Amino Acid ◽  
Signal Sequence ◽  
Residual Activity ◽  
Biodiesel Production ◽  
Molar Ratio ◽  
Subtilis Strain ◽  
Lipase Gene ◽  
E Coli

A lipase gene from sequencing genomic DNA of Bacillus subtilis strain I4 (I4-2) was cloned and expressed in E. coli. The deduced amino acid (aa) sequences for the lipase I4-2 was composed of 31-amino-acid signal sequence and a 181-amino-acid mature part, corresponding to a molecular mass (Mr) of 19.33 kDa. Based on the Mr and the protein sequence, the lipase I4-2 was belong to the lipase family1.4. The activity of the purified recombinant lipase I4-2 was apparently optimal at 45°C and pH 7.0. The enzyme was stable at 25°C, and more than 56% of the initial activity after incubation in buffer pH 7.0 for 120 min at 37°C. In addition, lipase I4-2 was mixed with methanol (50, v/v) for 30 min and residual activity was 45%. lipase I4-2 can catalyzed biodiesel production from soybean oil at methanol : soybean (molar ratio of methanol to oil = 3:1), 10% n-hexane, 3.5% water. The results of GC–MS analysis demonstrated that a 3-stepwise process resulted in a 98% conversion yield after 8 h of reaction at 37°C.

scholarly journals Establishment of electroporation protocol for Bacillus subtilis 1012 and WB800N

2015 ◽  
Vol 18 (2) ◽  
pp. 16-24
Author(s):  
Tien Thi My Pham ◽  
Trang Thi Phuong Phan
Keyword(s):  
Bacillus Subtilis ◽  
Recombinant Proteins ◽  
Culture Medium ◽  
Sampling Time ◽  
Extracellular Proteins ◽  
Bacillus Species ◽  
Subtilis Strain ◽  
Industrial Enzymes ◽  
Expression Host ◽  
Bacterial Genetic

Bacillus subtilis has been developed as an attractive expression host because of many advantages. For examples, it is nonpathogenic and allows secretion of functional extracellular proteins directly into the culture medium; about 60 % of industrial enzymes available produced by Bacillus species. To use B. subtilis strain for research and as host strain for expression of recombinant protein, bacterial genetic methods should be developed. Electroporation to transfer directly DNA into B. subtilis is one of the methods that draw a lot of attention of scientists. A problem encountered in the methods that draw a lot of attention of scientists. A problem encountered in the electroporation of DNA into B. subtilis is that an established protocol for one strain can hardly be used for another strain. B. subtilis 1012 and WB800N have recently been used as expression hosts for expression of recombinant proteins, but electroporation method has not been established. In this study, we use a pHT plasmid to establish an electroporation protocol for B. subtilis 1012 and WB800N. The influence of sampling time, concentration and time for incubating with lysozyme, voltage on the transformation was investigated to establish the protocol.

scholarly journals Two Tachykinin-Related Peptides with Antimicrobial Activity Isolated from Triatoma infestans Hemolymph

2020 ◽  
Vol 13 ◽  
pp. 117863612093363
Author(s):  
Laura Cristina Lima Diniz ◽  
Flávio Lopes Alves ◽  
Antonio Miranda ◽  
Pedro Ismael da Silva Junior
Keyword(s):  
Antimicrobial Activity ◽  
Amino Acid ◽  
Secondary Structure ◽  
High Performance ◽  
Micrococcus Luteus ◽  
Biological Activities ◽  
Biological Effects ◽  
Vero Cells ◽  
Reversed Phase ◽  
Triatoma Infestans

Antimicrobial peptides and proteins (AMPs) are molecules that can interact with microbial cells and lead to membrane disruption or intracellular molecule interactions and death. Several molecules with antimicrobial effects also present other biological activities. One such protein group representing the duplicity of activities is the tachykinin family. Tachykinins (TKs) form a family of neuropeptides in vertebrates with a consensus C-terminal region (F-X-G-Y-R-NH2). Invertebrate TKs and TK-related peptides (TKRPs) are subfamilies found in invertebrates that present high homology with TKs and have similar biological effects. Several of these molecules have already been described but reports of TKRP in Hemiptera species are limited. By analyzing the Triatoma infestans hemolymph by reversed-phase high-performance liquid chromatography, biological assays, and mass spectrometry, two antimicrobial molecules were isolated and identified as TKRPs, which we named as TRP1-TINF and TRP2-TINF (tachykinin-related peptides I and II from T. infestans). TRP1-TINF is a random secondary structure peptide with 9 amino acid residues. It is susceptible to aminopeptidases degradation and is active mainly against Micrococcus luteus (32 μM). TRP2-TINF is a 10-amino acid peptide with a 310 helix secondary structure and is susceptible to carboxypeptidases degradation. It has major antimicrobial activity against both Pseudomonas aeruginosa and Escherichia coli (45 μM). Neither molecule is toxic to human erythrocytes and both present minor toxicity toward Vero cells at a concentration of 1000 μM. As the first description of TKRPs with antimicrobial activity in T. infestans, this work contributes to the wider comprehension of the insects’ physiology and describes pharmacological relevant molecules.

scholarly journals Genome mining and UHPLC–QTOF–MS/MS illuminate the potential antimicrobial active compounds and specificity of biosynthetic gene clusters in Bacillus subtilis NCD-2

2020 ◽  
Author(s):  
Zhenhe Su ◽  
Xiuye Chen ◽  
Xiaomeng Liu ◽  
Qinggang Guo ◽  
Shezeng Li ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Gene Cluster ◽  
Biological Activities ◽  
Genome Mining ◽  
Gene Clusters ◽  
Integrative Approach ◽  
Amino Acid Sequence Similarity ◽  
Subtilis Strain ◽  
Biosynthetic Gene ◽  
Biosynthetic Gene Clusters

Abstract Background Bacillus subtilis strain NCD-2 is an excellent biocontrol agent against plant soil-borne diseases and shows broad-spectrum antifungal activities. This study aimed to explore some of the secondary metabolite biosynthetic gene clusters and related bioactive compounds in strain NCD-2. An integrative approach, which combined genome mining with structural identification technologies using ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight tandem mass spectrometry (UHPLC-MS/MS), was conducted to interpret the chemical origins of the significant biological activities in strain NCD-2. Results Genome mining revealed that strain NCD-2 contained nine gene clusters having predicted functions involving secondary metabolites with bioactive abilities. They encoded six known products including fengycin, surfactin, bacillaene, subtilosin, bacillibactin, bacilysin and three unknown products. Fengycin, surfactin, bacillaene and bacillibactin were successfully detected from the fermentation broth of strain NCD-2 by UHPLC-QTOF-MS/MS. Bacillaene, subtilosin, bacillibactin, and bacilysin related biosynthetic gene clusters showed 100% amino acid sequence similarity with B. velezensis strain FZB42，however, the biosynthetic gene clusters for surfactin and fengycin showed 83% and 92%, respectively. Further comparison of gene clusters encoding fengycin and surfactin revealed that strain NCD-2 had lost the fenC and fenD genes in the fengycin biosynthetic operon. Moreover, biosynthetic enzyme-related gene srfAB for surfactin had divided into two parts. Bioinformatics analysis predicted that FenE function in strain NCD-2 was same to that of FenE and FenC in strain FZB42, and FenA function in strain NCD-2 was same to that of FenA and FenD in strain FZB42. Five kinds of fengycin, with 26 homologs, and surfactin, with 4 homologs, were detected from strain NCD-2. To the best of our knowledge, this is the first report of a non-typical and unique gene cluster related to fengycin synthesis. Conclusions It was found that there were many gene clusters encoding antimicrobial compounds in the genome of strain NCD-2, and the fengycin synthetic gene cluster might be unique by using genome mining and UHPLC–QTOF–MS/MS. The production of fengycin, surfactin, bacillaene and bacillibactin might explain the biological activities of strain NCD-2.

scholarly journals Purification of surfactin compounds produced by a Bacillus subtilis strain

2021 ◽  
Vol 64 (2) ◽  
pp. 121-128
Author(s):  
Attila Bartal ◽  
Henriett Hunkár ◽  
Gábor Endre ◽  
Mónika Vörös ◽  
Csaba Vágvölgyi ◽  
...  
Keyword(s):  
Crude Extract ◽  
Biological Activities ◽  
Flash Chromatography ◽  
Bacillus Species ◽  
Subtilis Strain ◽  
Preparative Hplc ◽  
Reverse Phase Hplc ◽  
Hplc Purification ◽  
Fatty Acid Chain

Surfactins are lipopeptide-type biosurfactants produced mainly by Bacillus species containing a peptide loop of seven amino acids and a hydrophobic fatty acid chain. These molecules exhibit various biological activities; therefore, their therapeutic and environmental applications are in the focus worldwide. In our work, a multi-step purification and separation process was developed to isolate surfactins from the ferment broth of B. subtilis SZMC 6179J strain. The process incorporates normal phase flash chromatography for pre-purifying the crude extract and two consecutive reverse phase HPLC separations for the isolation of the various surfactin molecules. The determination of the relative amounts of lipopeptides both in the crude extract and in each fraction of every separation step were carried out by HPLC-HESI-MS examinations. The ratio of surfactins in the crude extract was 21.35%, but after the preparative flash chromatographic separation the relative amount of surfactins was observed to be 30.44%. The preparative HPLC purification step resulted 85.39% purity of the surfactins. Nine different surfactin variants were isolated and identified from the fractions of this final semi-preparative HPLC purification, out of which three compounds were completely purified, and three others were detected in relative amounts of more than 95% in some fractions.

Current Advances in the Biological Activity of Polysaccharides in Dendrobium with Intriguing Therapeutic Potential

2018 ◽  
Vol 25 (14) ◽  
pp. 1663-1681 ◽  
Author(s):  
Chun-Ting Lee ◽  
Heng-Chun Kuo ◽  
Yung-Hsiang Chen ◽  
Ming-Yen Tsai
Keyword(s):  
Biological Activity ◽  
Therapeutic Potential ◽  
Biological Activities ◽  
Biological Effects ◽  
Herbal Medicines ◽  
Research Field ◽  
Protective Effects ◽  
Pharmacological Effects ◽  
The Family ◽  
Anti Tumor Activity

The polysaccharides in many plants are attracting worldwide attention because of their biological activities and medical properties, such as anti-viral, anti-oxidative, antichronic inflammation, anti-hypertensive, immunomodulation, and neuron-protective effects, as well as anti-tumor activity. Denodrobium species, a genus of the family orchidaceae, have been used as herbal medicines for hundreds of years in China due to their pharmacological effects. These effects include nourishing the Yin, supplementing the stomach, increasing body fluids, and clearing heat. Recently, numerous researchers have investigated possible active compounds in Denodrobium species, such as lectins, phenanthrenes, alkaloids, trigonopol A, and polysaccharides. Unlike those of other plants, the biological effects of polysaccharides in Dendrobium are a novel research field. In this review, we focus on these novel findings to give readers an overall picture of the intriguing therapeutic potential of polysaccharides in Dendrobium, especially those of the four commonly-used Denodrobium species: D. huoshanense, D. offininale, D. nobile, and D. chrysotoxum.

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