Production of lipopeptide biosurfactant by Bacillus subtilis GY19 and its application as oil-contaminated surface cleaning agent

ABSTRACTMulticellular communities produced byBacillus subtiliscan adopt sliding or swarming to translocate over surfaces. While sliding is a flagellum-independent motility produced by the expansive forces in a growing colony, swarming requires flagellar functionality and is characterized by the appearance of hyperflagellated swarm cells that associate in bundles or rafts during movement. Previous work has shown that swarming by undomesticatedB. subtilisstrains requiresswrA, a gene that upregulates the expression of flagellar genes and increases swimming motility, and surfactin, a lipopeptide biosurfactant that also facilitates sliding. Through an analysis ofswrA+andswrAmutant laboratory strains with or without a mutation insfp(a gene involved in surfactin production), we show that bothswrAand surfactin upregulate the transcription of the flagellin gene and increase bacterial swimming. Surfactin also allows the nonswarmingswrAmutant strain to efficiently colonize moist surfaces by sliding. Finally, we reconfirm the essential role ofswrAin swarming and show that surfactin, which increases surface wettability, allowsswrA+strains to produce swarm cells on media at low humidity.

Download Full-text

Characterization of Lipopeptides Biosurfactants Produced by a Newly Isolated Strain Bacillus Subtilis ZNI5: Potential Environmental Application

10.21203/rs.3.rs-308797/v1 ◽

2021 ◽

Author(s):

Mnif Inès ◽

Hayfa Rajhi ◽

Amir Bouallegue ◽

Nour Trabelsi ◽

Dhouha Ghribi

Keyword(s):

Surface Tension ◽

Bacillus Subtilis ◽

High Performance ◽

Surface Active Agent ◽

Active Agent ◽

Industrial Applications ◽

Exchange Chromatography ◽

Environmental Application ◽

Molecular Weights ◽

Lipopeptide Biosurfactant

Abstract Strain ZNI5, isolated from a hydrocarbon contaminated soil and identified as Bacillus subtilis after 16s rDNA sequence, grew and produced lipopeptides biosurfactants when cultured on glucose based media. After purification by anionic exchange chromatography and identification Reverse Phase High Performance Liquid Chromatography-Mass Spectrometry, the biosurfactant produced by ZNI5 were determined to be cyclic lipopeptides homologues. Four families of lipopeptides were identified by HPLC-MS analysis. They belongs to surfactin isoforms with molecular weights of 979, 993, 1007, 1021 and 1035 Da; iturin isoforms with molecular weights of 1028, 1042 and 1056 Da; Licheniformin with molecular weight of 1410 and newly identified isoforms named Inesfactin with molecular weights of 973 and 987 Da. Functional properties of the ZNI5 biosurfactant were studied. It was characterized as a powerful surface-active agent that decreases the surface tension of water from 72 mN/m to about 32 mN/m with a CMC value of 350 mg/L more efficient than chemical surfactants (Triton X100; CTAB and SDS). It has the capacity to disperse oil to about 80 mm at a concentration of 800 mg/L showing close efficiencies to the listed chemical surfactants. In addition, by studying the surface tension decrease capacity and the oil displacement activity, ZNI5 lipopeptide biosurfactant showed great thermal, pH and salts activity and stability enabling its use in the bioremediation fields and for diverse industrial applications.

Download Full-text

Production and applications of lipopeptide biosurfactant for bioremediation and oil recovery by Bacillus subtilis CN2

Biochemical Engineering Journal ◽

10.1016/j.bej.2015.05.007 ◽

2015 ◽

Vol 101 ◽

pp. 168-178 ◽

Cited By ~ 79

Author(s):

Fisseha Andualem Bezza ◽

Evans M. Nkhalambayausi Chirwa

Keyword(s):

Bacillus Subtilis ◽

Oil Recovery ◽

Lipopeptide Biosurfactant

Download Full-text

Gene yerP, Involved in Surfactin Self-Resistance in Bacillus subtilis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.12.3566-3573.2001 ◽

2001 ◽

Vol 45 (12) ◽

pp. 3566-3573 ◽

Cited By ~ 43

Author(s):

Kenji Tsuge ◽

Yuichiro Ohata ◽

Makoto Shoda

Keyword(s):

Bacillus Subtilis ◽

Cell Division ◽

Proton Motive Force ◽

Phosphopantetheinyl Transferase ◽

Logarithmic Growth Phase ◽

Gram Negative ◽

Cyclic Lipopeptide ◽

Lipopeptide Biosurfactant ◽

Surfactin Production ◽

Logarithmic Growth

ABSTRACT Surfactin is a cyclic lipopeptide biosurfactant. Transposon mutagenesis was performed in Bacillus subtilis strain 168, and a surfactin-susceptible mutant, strain 801, was isolated. Analysis of the region of insertion revealed that yerPwas the determinant of surfactin self-resistance. YerP had homology with the resistance, nodulation, and cell division (RND) family proton motive force-dependent efflux pumps only characterized in gram-negative strains. The yerP-deficient strain 802, in which the internal region of the yerP gene of B.subtilis strain 168 was deleted, showed susceptibility to acriflavine and ethidium bromide. When strain 802 was converted to a surfactin producer by introducing a functional sfp which encodes a 4′-phosphopantetheinyl transferase and is mutated inB. subtilis strain 168, thisyerP-deficient strain produced surfactin, although surfactin production was significantly reduced. The expression ofyerP was at its maximum at the end of the logarithmic growth phase and was not induced by surfactin. yerP is the first RND-like gene characterized in gram-positive strains and is supposed to be involved in the efflux of surfactin.

Download Full-text

Antagonistic activity of lipopeptide-biosurfactant producing Bacillus subtilis AKP, against Colletotrichum capsici, the causal organism of anthracnose disease of chilli

Biocatalysis and Agricultural Biotechnology ◽

10.1016/j.bcab.2021.102133 ◽

2021 ◽

Vol 36 ◽

pp. 102133

Author(s):

Amrit Kumar ◽

Jintu Rabha ◽

Dhruva Kumar Jha

Keyword(s):

Bacillus Subtilis ◽

Antagonistic Activity ◽

Causal Organism ◽

Anthracnose Disease ◽

Colletotrichum Capsici ◽

Lipopeptide Biosurfactant

Download Full-text

Impact of the Purification Process on the Spray-Drying Performances of the Three Families of Lipopeptide Biosurfactant Produced by Bacillus subtilis

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2021.815337 ◽

2021 ◽

Vol 9 ◽

Author(s):

Antoine Vassaux ◽

Marie Rannou ◽

Soline Peers ◽

Théo Daboudet ◽

Philippe Jacques ◽

...

Keyword(s):

Bacillus Subtilis ◽

Spray Drying ◽

Freeze Drying ◽

Industrial Applications ◽

Purification Process ◽

Purification Step ◽

Wide Range ◽

Lipopeptide Biosurfactant ◽

The Impact ◽

Spray Dried

Lipopeptides produced by Bacillus subtilis display many activities (surfactant, antimicrobial, and antitumoral), which make them interesting compounds with a wide range of applications. During the past years, several processes have been developed to enable their production and purification with suitable yield and purity. The already implemented processes mainly end with a critical drying step, which is currently achieved by freeze-drying. In this study, the possibility to replace this freeze-drying step with a spray-drying one, more suited to industrial applications, was analyzed. After evaluating their thermal resistance, we have developed a spray-drying methodology applicable for the three lipopeptides families produced by B. subtilis, i.e., surfactin, mycosubtilin (iturin family), and plipastatin (fengycin family). For each lipopeptide, the spray-drying procedure was applied at three steps of the purification process by ultrafiltration (supernatant, diafiltered solution, and pre-purified fraction). The analysis of the activities of each spray-dried lipopeptide showed that this drying method is not decreasing its antimicrobial and biosurfactant properties. The methodology developed in this study enabled for the first time the spray-drying of surfactin, without adjuvants’ addition and regardless of the purification step considered. In the case of fengycin and mycosubtilin, only diafiltered solution and purified fraction could be successfully spray-dried without the addition of adjuvant. Maltodextrin addition was also investigated as the solution for the direct drying of supernatant. As expected, the performances of the spray-drying step and the purity of the powder obtained are highly related to the purification step at which the product was dried. Interestingly, the impact of mycosubtilin concentration on spray-drying yield was also evidenced.

Download Full-text