Production Of Glycolipid Biosurfactant Research Articles

Production and characterization of glycolipid biosurfactant from Streptomyces enissocaesilis HRB1 and its evaluation for biomedical and bioremediation applications

AbstractThe aim of this investigation is to produce and characterize biosurfactant from Streptomyces sp. HRB1 and to evaluate its biomedical and bioremediation potential. Biosurfactant producing property of Streptomyces sp. HRB1 isolated from petroleum contaminated soil was confirmed by hemolytic and oil spread assays. Based on the results of FT‐IR spectral and GC–MS analysis, the biosurfactant was confirmed as glycolipid type. Biosurfactant from Streptomyces sp. HRB1 exhibited 71% inhibition against Pseudomonas aeruginosa biofilm formation, 77.33% quorum sensing inhibition property against Chromobacterium violeceum MTCC 2656, more than 80% inhibition in antioxidant assays namely, DPPH, ABTS, and metal chelation, promising anti‐proliferative activity against leukemia and myeloma cells with low IC50 values, 96% decolorization of malachite green within 48 h of reaction time, and minimal toxicity against normal cell lines in dose‐dependent manner. The taxonomic position of the potential strain HRB1 was further confirmed as Streptomyces enissocaesilis HRB1 based on their phenotypic and molecular characteristics. To conclude, Streptomyces enissocaesilis HRB1 isolated from petroleum‐contaminated soil is a promising source for low‐cost production of glycolipid biosurfactant with potential biomedical and environmental applications such as antiphytofungal, antibiofilm, anti‐quorum sensing, antioxidant, anticancer, and dye degradation properties.

Antinociceptive and anti-inflammatory activity of α-d-mannan from Pseudozyma sp.

The online version contains supplementary material available at 10.1007/s13205-020-02635-1.

Production of glycolipid biosurfactant during crude oil degradation by the novel indigenous isolated Achromobacter kerstersii LMG3441.

This study aimed to find biosurfactant producing and crude oil-degrading bacteria able to decontaminate crude oil from wastewater. The bacteria that were isolated from contaminated sites in an oil refinery plant in Isfahan, Iran, were identified by 16S rDNA sequencing as Achromobacter kerstersii strain LMG3441, Klebsiella pneumonia strain SKBA6, and Klebsiella variicola strain SKV2. According to the results obtained from different tests for the production of biosurfactant among three strains, only Achromobacter kerstersii strain LMG3441 was selected for further study. The pattern of residual hydrocarbons was analyzed by high-resolution gas chromatography-mass spectrometry (GC-MS). This novel and indigenous strain was capable of producing the highest amount of a glycolipid biosurfactant (7.81 g/L) in MSM (mineral salt medium) with 1% (v/v) crude oil as the only source of carbon and energy. The compound showed high surface activation capacity with reduction of surface tension from 40 mN m-1 in the control to 23.3 mN m-1 by the bacterium. The results of GC-MS for assessment of residual hydrocarbons in the MSM and comparison with crude oil as a control showed that 53% of the hydrocarbons in the crude oil were consumed by this novel strain.

Valorization of xylose-rich hydrolysate from rice straw, an agroresidue, through biosurfactant production by the soil bacterium Serratia nematodiphila

Biosurfactants, amphiphilic compounds that reduce interfacial tension in oil-aqueous mixtures, are used in the petroleum, pharmaceutical, food, and agriculture industries. Fermentative production of biosurfactants requires expensive sugar or lipid substrates. Lignocellulosic biomass is a relatively cheap and abundant agricultural residue that can be used as an alternative substrate. Currently, several million tonnes of rice and wheat straw are generated globally as agricultural residues, most of which is disposed by open-field burning thereby leading to severe environmental pollution. This study aimed to produce biosurfactants in xylose-rich hydrolysates generated from rice straw. The hydrolysate is also a byproduct of 2G biofuel processes that often goes underutilized. A soil bacterium capable of growing and producing biosurfactants in rice straw hydrolysates, which typically contain growth-inhibitory compounds such as furfural and hydroxymethyl furfural, was isolated. Interestingly, the organism, identified as Serratia nematodiphila, exhibited higher glycolipid formation (4.5 ± 0.6 gL−1) in xylose-rich hydrolysate than in glucose-rich enzymatic hydrolysate (3.1 ± 0.2 gL−1) despite the higher bacterial cell density observed with the latter. The biosurfactants were thermostable and possessed promising emulsifying property and anti-microbial activity against bacteria and yeast. Further optimization of C:N resulted in a 2.8-fold increase in glycolipid production from xylose-rich hydrolysates. This study demonstrates the production of glycolipid biosurfactants from lignocellulosic biomass, a low-cost substrate and offers a plausible strategy for the management of these residues. Further, it also provides insights into the generation of additional high-value compounds in a bioethanol biorefinery to improve its commercial feasibility.

A Review on Biosurfactants and its Environmental Applications

Biosurfactants are widely known as surface active agents of biological origin. Glycolipid classes of biosurfactants has high important in the biotechnological arena. Pseudomonas aeruginosa, Bacillus subtilis, and Candida sp., are important classes of microorganism and highly investigated for the production of glycolipid biosurfactants. Commercially, microbial biosurfactants are more advantageous than chemical based biosurfactants due to their biodegradability, renewability and functionality maintenance under extreme operating conditions. In the oil spill areas hydrocarbon degrading microorganisms (isolated) were proven to produce enormous amount of biosurfactant than expected. This is due to the regulation of all the genomes in the synthesis of lipid metabolism. Currently, biosurfactants play vital role in oil and petroleum industries for emulsification in both recovery and removal process from the site of pollution. In addition, it has some role in the heavy metal removal of metallurgical industries. In the present paper, we have given the overview on screening of biosurfactant producing microorganisms, production methods and factors influencing the production of biosurfactants are investigated. Various analytical techniques used for processing of crude metabolites are also presented. Thus from this review, can easily understood the role of biosurfactant in the environmental cleaning.

Glycolipid biosurfactants: main properties and potential applications in agriculture and food industry.

Glycolipids, consisting of a carbohydrate moiety linked to fatty acids, are microbial surface active compounds produced by various microorganisms. They are characterized by high structural diversity and have the ability to decrease the surface and interfacial tension at the surface and interface, respectively. Rhamnolipids, trehalolipids, mannosylerythritol lipids and cellobiose lipids are among the most popular glycolipids. They have received much practical attention as biopesticides for controlling plant diseases and protecting stored products. As a result of their antifungal activity towards phytopathogenic fungi and larvicidal and mosquitocidal potencies, glycolipid biosurfactants permit the preservation of plants and plant crops from pest invasion. Also, as a result of their emulsifying and antibacterial activities, glycolipids have great potential as food additives and food preservatives. Furthermore, the valorization of food byproducts via the production of glycolipid biosurfactant has received much attention because it permits the bioconversion of byproducts on valuable compounds and decreases the cost of production. Generally, the use of glycolipids in many fields requires their retention from fermentation media. Accordingly, different strategies have been developed to extract and purify glycolipids. © 2016 Society of Chemical Industry.

English

Biosurfactants are amphiphilic compounds produced by bacteria and fungi to reduce surface and interfacial tension. This work was designed to produce biosurfactants from the fermentation of submerge cashew bagasse (Anacardium occidentale) using a microorganism Pseudomonas aeruginosa. The proximate components of the biosurfactant was determined. Results show that yellow cashew fruit bagasse contains lipid (11.34 ± 0.16%), protein (26.67 ± 0.66%), carbohydrate (49.37 ± 0.60%), moisture (5.78 ± 0.17%), ash (2.70 ± 0.04%) and fibre (2.86 ± 0.09%). Biosurfactants activity was characterized based on emulsification index and oil displacement capacity. The yield of biosurfactants was, 0.71 g; when only cashew was used; 0.93 g for cashew and glucose and nutrient broth 0.12 g respectively. The preliminary biochemical characterization revealed that the biosurfactants contained carbohydrates and lipids designated as glycolipids. The biosurfactants showed antimicrobial activity against a range of Gram positive and Gram negative bacteria strains with diameters of zone and growth inhibition: Escherichia coli, 38.70 ± 1.30 mm; Staphylococcus aureus, 38.00 ± 2.94 mm; Klebsiella, 31.00 ± 2.20 mm and Bacillus cereus, 28.70 ± 1.70 mm. These results suggest that cashew fruits bagasse serve as cheap carbon source for the production of glycolipid biosurfactants with useful industrial applications. Key words: Biosurfactants, fruit bagasse, yellow cashew, characterization and Pseudomonas aeruginosa.

Production of glycolipid biosurfactants, mannosylerythritol lipids, from pentoses and d-glucose/d-xylose mixtures by Pseudozyma yeast strains

The pentose-assimilating capacity of yeasts from the genus Pseudozyma, P. antarctica PYCC 5048T, P. aphidis PYCC 5535T and P. rugulosa PYCC 5537T, was exploited towards the production of mannosylerythritol lipids (MEL), a glycolipid with biosurfactant properties. The three strains tested were able to grow on d-xylose and l-arabinose with similar maximum specific growth rates to those estimated on d-glucose (around 0.2h−1). The highest MEL titres (4.8–5.4g/l) and yields (0.11–0.14g/g) from d-xylose were found in P. antarctica PYCC 5048T, which presented similar values to those estimated on d-glucose and on d-xylose/d-glucose mixtures. P. rugulosa PYCC 5537T showed a pattern of sugar conversion into MEL similar to P. antarctica, but at 40% lower titres. P. aphidis PYCC 5535T presented lower MEL titres from d-xylose (1.2g/l) than from d-glucose (3.4g/l). Nitrate supply increased sugar consumption rate and, when accompanied by d-glucose or d-xylose feeding, additional biomass production. In this case, sugar was completely consumed before sugar feeding at day 7, but not when feeding is performed at day 4. Higher MEL titres where obtained for the later condition reaching 7.3g/l and 5.8g/l, in fed-batch cultures with glucose and xylose, respectively.

Optimization, production and characterization of glycolipid biosurfactant from the marine actinobacterium, Streptomyces sp. MAB36

A potential glycolipid biosurfactant producer Streptomyces sp. MAB36 was isolated from marine sediment samples. Medium composition and culture conditions for the glycolipid biosurfactant production by Streptomyces sp. MAB36 were optimized, using two statistical methods: Plackett-Burman design was applied to find out the key ingredients and conditions for the best yield of glycolipid biosurfactant production and central composite design was used to optimize the concentration of the four significant variables, starch, casein, crude oil and incubation time. Fructose and yeast extract were the best carbon and nitrogen sources for the production of the glycolipid biosurfactant. Biochemical characterizations including FTIR and MS studies suggested the glycolipid nature of the biosurfactant. The isolated glycolipid biosurfactant reduced the surface tension of water from 73.2 to 32.4mN/m. The purified glycolipid biosurfactant showed critical micelle concentrations of 36mg/l. The glycolipid biosurfactant was effective at very low concentrations over a wide range of temperature, pH, and NaCl concentration. The purified glycolipid biosurfactant showed strong antimicrobial activity. Thus, the strain Streptomyces sp. MAB36 has proved to be a potential source of glycolipid biosurfactant that could be used for the bioremediation processes in the marine environment.

Crude biosurfactant from thermophilic Alcaligenes faecalis: Feasibility in petro-spill bioremediation

The thermophilic bacterium Alcaligenes faecalis isolated from the crude oil contaminated soil of Upper Assam, India. The isolated bacterium was first screened for the ability to produce biosurfactant. The strain growing at 42 °C could produce higher amount of biosurfactant in medium supplemented with 2% (v/v) diesel as sole source of carbon and energy. Biochemical characterizations including FT-IR and MS studies suggested the biosurfactant to be glycolipid. Tensiometric studies revealed that the biosurfactant produced by the bacterial strain could decrease the surface tension ( σ) at air–water interface from 71.6 to 32.3 mNm −1 after 96 h of growth on hydrocarbon and possessed a low critical micelle concentration (CMC) value of approximately 38 mgl −1, indicating high surface activity. The culture supernatant containing the biosurfactant was found to be functionally stable at varying pH (2–12), temperature (100 and 121 °C) and salinity (1–6% NaCl, w/v) conditions. Both the culture broth and the cell free supernatant exhibited high emulsifying activity against the different hydrocarbons and the crude oil components. The increase in cell surface hydrophobicity and glycolipid production by the strain suggested the existence of biosurfactant enhanced interfacial uptake of the hydrocarbons. Moreover, the partially purified biosurfactant exhibited antimicrobial activity by inhibiting the growth of several bacterial and fungal species. The strain represented a new class of biosurfactant producers and could be a potential candidate for the production of glycolipid biosurfactant which could be useful in a variety of biotechnological and industrial processes, particularly in the oil industry.

Yeast extract stimulates production of glycolipid biosurfactants, mannosylerythritol lipids, by Pseudozyma hubeiensis SY62

We improved the culture conditions for a biosurfactant producing yeast, Pseudozyma hubeiensis SY62. We found that yeast extract greatly stimulates MEL production. Furthermore, we demonstrated a highly efficient production of MELs in the improved medium by fed-batch cultivation. The final concentration of MELs reached 129 ± 8.2g/l for one week.

Production of Glycolipid Biosurfactants, Mannosylerythritol Lipids, Using Sucrose by Fungal and Yeast Strains, and Their Interfacial Properties

Glycolipid biosurfactants, mannosylerythritol lipids (MELs), were produced from glucose and sucrose without vegetable oils. Pseudozyma antarctica JCM 10317, Ustilago maydis NBRC 5346, U. scitaminea NBRC 32730, and P. siamensis CBS 9960 produced mainly MEL-A, MEL-A, MEL-B, and MEL-C respectively. The sucrose-derived MELs showed excellent interfacial properties: low critical micelle concentration as well as that of oil-derived MELs.

Production of glycolipid biosurfactants by basidiomycetous yeasts

BSs (biosurfactants) produced by various micro-organisms show unique properties (e.g. mild production conditions, lower toxicity, higher biodegradability and environmental compatibility) compared with chemically synthesized surfactants. The numerous advantages of BSs have prompted applications not only in the food, cosmetic and pharmaceutical industries but also in environmental protection and energy-saving technology. Among BSs, glycolipid types are the most promising, owing to their high productivity from renewable resources and versatile biochemical properties. MELs (mannosylerythritol lipids), which are glycolipid BSs abundantly produced by basidiomycetous yeasts such as strains of Pseudozyma, exhibit not only excellent interfacial properties, but also remarkable differentiation-inducing activities against human leukaemia cells. MELs also show high binding affinity towards different immunoglobulins and lectins. Recently, a cationic liposome bearing MEL has been demonstrated to increase dramatically the efficiency of gene transfection into mammalian cells. These features of BSs should broaden their application in new advanced technologies. In the present review the current status of research and development on glycolipid BSs, especially their production by Pseudozyma yeasts, is described.

Production of glycolipid biosurfactants, mannosylerythritol lipids, by Pseudozyma siamensis CBS 9960 and their interfacial properties

The search for a novel producer of glycolipid biosurfactants, mannosylerythritol lipids (MELs), was undertaken on the basis of the analysis of ribosomal DNA sequences of yeast strains of the genus Pseudozyma. In the course of the investigation, Pseudozyma siamensis CBS 9960, which is closely related to Pseudozyma shanxiensis, a known MEL-C producer but with a different morphology, was found to accumulate a large amount of glycolipids. On thin layer chromatography, the extracellular glycolipids showed nearly the same spots as those of the MELs produced by P. shanxiensis. However, the result of high-performance liquid chromatography analysis revealed that the present strain has a much higher glycolipid production yield than P. shanxiensis. From the structural characterization by (1)H and (13)C NMR, the major glycolipid (more than 84% of the total) was identified as a mixture of 4-O-[(2',4'-di-O-acetyl-3'-O-alka(e)noyl)-beta-D-mannopyranosyl]-D-erythritol and 4-O-[(4'-O-acetyl-3'-O-alka(e)noyl-2'-O-butanoyl)-beta-D-mannopyranosyl]-D-erythritol, both of which are types of MEL-C. The present MEL-C possessed a short-chain acid (C(2) or C(4)) at the C-2' position and a long-chain acid (C(16)) at the C-3' position of the mannose moiety, and thus, the hydrophobic part was considerably different from that of conventional MELs, which mainly possess two medium-chain acids (C(10)) at the C-2' and C-3' positions. Under optimal growth conditions with safflower oil in a shake culture, the total amount of MELs reached approximately 19 g/l after 9 d at 25 degrees C. We further investigated the interfacial properties of the present MEL-C, considering its unique hydrophobic structure. The observed critical micelle concentration (CMC) and the surface tension at the CMC of the MEL were 4.5 x 10(-6) M and 30.7 mN/m, respectively. In addition, on a water penetration scan, the MEL efficiently formed the liquid crystal phases such as hexagonal (H) and lamella (L(a)) at a wide range of concentrations. These results demonstrated that the newly identified MEL-C produced by P. siamensis exhibits not only high surface activity but also excellent self-assembling properties, and should facilitate the development of promising yeast biosurfactants.

Genetic Regulations of the Biosynthesis of Microbial Surfactants: An Overview

Microbial biosurfactants are surface active metabolites synthesized by microbes growing on a variety of substrates. In spite of having great potential for commercial, therapeutic and environmental applications, industrial level production has not been realized for their low yields and productivities. One vital factor determining their biosynthesis is the genetic makeup of the producer organisms. Studies on molecular genetics and biochemistry of the synthesis of several biosurfactants have revealed the operons, the enzymes and the metabolic pathways required for their extracellular production. Surfactin, a cyclic lipopeptide biosurfactant is a potent antimicrobial agent and is produced as a result of non-ribosomal biosynthesis catalyzed by a large multienzyme peptide synthetase complex called the surfactin synthetase. Pathways for the synthesis of other lipopeptides such as iturin, lichenysin and arthrofactin are also mediated by similar enzyme complexes. These non-ribosomal peptide synthetases (NRPSs) responsible for lipopeptide biosynthesis display a high degree of structural similarity among themselves even from distant microbial species. Plasmid-encoded- rhlA, B, R and I genes of rhl quorum sensing system are required for production of glycolipid biosurfactants by Pseudomonas species. Molecular genetics of biosynthesis of alasan and emulsan by Acinetobacter species and of the fungal biosurfactants such as mannosylerythritol lipids (MEL) and hydrophobins have been deciphered. However, limited genetic information is available about biosynthesis of other biosurfactants such as viscosin, amphisin and synthesized counterparts. As these molecules have emerged as potential agents in many industrial and environmental processes as well as in biomedical and therapeutic applications, it is essential to make them cost competitive. Genetically engineered hyper producing organisms giving high yields can bring the real breakthrough in the production process. This is possible only if the genetics of the microbial surfactant production is known in details. It is therefore desirable that the future research on biosurfactants be focused on the development and use of hyperproducers. The detailed knowledge of the genetics of microbial surfactant production should be used to produce organisms giving higher production with better product characteristics. With a better knowledge of the genes involved in this process, biosurfactant production can be realized in non-pathogenic industrial strains. The knowledge of expression of genes of a particular biosurfactant producer in a particular habitat will also throw light upon substrate dependence of production and preference for a particular substrate. Detailed description of the genetics of production of the newly identified biosurfactants like flavolipids, tensin and lokisin is not available. Efforts should be made by investigators to develop high yielding strains of microorganisms producing strong biosurfactants such as arthrofactin and lichenysin. A few marine biosurfactants have been discovered (Kalinovskaya et al. 2004) and there is high possibility of finding many other novel surface active compounds from the marine sources. Many of these are expected to possess interesting properties as pharmaceuticals and biomedical agents. The knowledge of molecular genetics of microbial surfactant production and its subsequent use to produce hyperproducers will determine the fate of biosurfactant industry.

バイオサーファクタントの生産と機能利用に関する研究

バイオサーファクタントの生産と機能利用に関する研究