Consortium of lipophilic bacterial strains for biodegrading palm oil

Abstract

The invention relates to the field of biotechnology, and more particularly to a consortium of lipophilic bacterial strains for biodegrading palm oil, consisting of Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-3071D in a 1:1 ratio. The claimed consortium of lipophilic bacterial strains can be use for breaking down palm oil mill effluent. The aim of the invention is to produce a novel consortium of lipophilic bacterial strains for biodegrading palm oil, characterized in that two bacterial strains capable of active growth in an environment containing palm oil as the sole source of organic matter are combined in a single association.

Description

[0001] Consortium of lipophilic bacterial strains for palm oil biodegradation

[0002] Field of technology

[0003] The invention relates to biotechnology and microbiology, specifically to a consortium of lipophilic bacterial strains Pseudomonas protegens VKM B-3844D and Gordonia paraffinnivorans VKM Ac-307 ID for the biodegradation of palm oil. This consortium of lipophilic bacterial strains can be used to degrade liquid waste from palm oil production.

[0004] Prior art

[0005] Liquid waste from palm oil production is characterized by a high content of unprocessed oil and other pollutants. Direct discharge of such effluents into the environment without proper treatment can cause serious environmental problems [Dominic D., Baidurah S. Recent developments in biological processing technology for palm oil mill effluent treatment - A review / / Biology - 2022. - Vol. 11. - P. 525]. Bacteria play a significant role in the biogeochemical cycles of natural and anthropogenic ecosystems. In river ecosystems, bacteria intensively colonize silt sediments; bottom sediments can be a source of metabolically diverse microorganisms, including those promising for industrial biotechnology [Araya R., Tani K., Takagi T., Yamaguchi N., Nasu M. Bacterial activity and community composition in stream water and biofilm from an urban river determined by fluorescent in situ hybridization and DGGE analysis. FEMS Microbiol. Ecol. 2003;43(l):ll l- 119. DOI 10.1111 / j.l5746941.2003.tb01050.x]. The search for effective microorganisms-destructors and the development of biotechnologies based on them has great potential for the development of industrial technologies for the biodegradation of palm oil.

[0006] The results of isolating lipophilic microorganisms from samples of liquid waste from the Surabaya treatment plant in Indonesia were previously published. Based on phylogenetic analysis, determination of lipolytic activity and ability to grow on a medium with palm oil, the Pseudomonas protegens strain was recognized as promising for further research [Gerasimchuk A. L., Ivasenko D. A., Trifonov A. A., Topilina Yu. S., Sysoeva A. N., Frank Yu. A. Search and isolation of lipophilic bacteria for the utilization of palm oil production waste / / In the book: BIOTECHNOLOGY: STATE AND DEVELOPMENT PROSPECTS. Proceedings of the international congress. Moscow, 2023. pp. 114-116]. Gordonia paraffinnivorans, an actinomycete with the ability to degrade hydrocarbons, was isolated from an oil-producing well in Daqing, China [Xue Y, Sun X, Zhou P, Liu R, Liang F, Ma Y. Gordonia paraffinnivorans sp. nov., a hydrocarbon-degrading actinomycete isolated from an oil-producing well. Int J Syst Evol Microbiol.2003 Sep;53(Pt 5):1643-1646. doi: 10.1099 / ijs.0.02605-0. PMID: 13130063]. No previously published data on the study of lipolytic activity of Gordonia paraffinnivorans in relation to palm oil were found.

[0007] Disclosure of invention

[0008] The objective of this invention is to obtain a new consortium of lipophilic bacterial strains for the biodegradation of palm oil, the distinctive feature of which is the combination in one association of two bacterial strains capable of active growth in a medium with palm oil as the only source of organic matter.

[0009] To solve the problem, a consortium of lipophilic bacterial strains for the biodegradation of palm oil was designed, consisting of Pseudomonas protegens VKM B-3844D and Gordonia paraflnivorans VKM Ac-307 ID in a 1:1 ratio.

[0010] Information on the deposition of strains

[0011] The strains were deposited in the All-Russian Collection of Microorganisms “FEDERAL RESEARCH CENTER “PUSHCHINSK SCIENTIFIC CENTER FOR BIOLOGICAL RESEARCH OF THE RUSSIAN ACADEMY OF SCIENCES”, SKRYABIN INSTITUTE OF BIOCHEMISTRY AND PHYSIOLOGY OF MICROORGANISMS OF THE RUSSIAN ACADEMY OF SCIENCES (IBPMMS RAS) (142290, Russia, Moscow region, Pushchino, pr. Nauki, 5) under registration numbers VKM B-3844D and VKM AC-3071D.

[0012] Pseudomonas protegens strain VKM B-3844D

[0013] Pseudomonas protegens strain VKM B-3844D was isolated from liquid waste samples from a wastewater treatment plant in Surabaya, Indonesia. The liquid waste was used as an inoculum for dilution culture on a synthetic agar medium supplemented with palm oil and carboxymethylcellulose at a final concentration of 1%, and on Plate Count Agar (PCA) rich nutrient medium.

[0014] Cultural, morphological, physiological, and biochemical characteristics: Colonies are light beige and opaque. Cells are rod-shaped, 2–4 µm long. No spores were detected. Cultivation temperature: 28°C; culturing time: 1–2 days.

[0015] Cultivation and storage

[0016] Pseudomonas protegens VKM B-3844D grows on media such as fish meal hydrolysate (FMH), Plate Count Agar (PCA), tributyrin agar (TBA), and mineral medium with palm oil at a final concentration of 1%. When grown on TBA, the strain forms hydrolysis zones around colonies of 1-2 mm, indicating the production of extracellular lipolytic enzymes. Growth on rich nutrient media takes 1-2 days at a temperature of 4-35°C, with an optimum of 28-32°C. The strain is capable of growth on mineral medium with palm oil (1%); growth takes 2-3 days at a temperature of 15-35°C, with an optimum of 28°C.

[0017] The Pseudomonas protegens strain VKM B-3844D is maintained in laboratory conditions by periodic subcultures once every 2-3 months on Plate Count Agar (PCA) agar medium consisting of: yeast extract 2.5 g / l; NaCl 5.0 g / l; D-glucose 1.0 g / l; dry enzymatic peptone 5.0 g / l; microbiological agar 15.0 g / l, distilled water to 1.0 l.

[0018] The Pseudomonas protegens strain VKM B-3844D can also be stored at a positive temperature of 4°C in a lyophilized state using natural skim milk as a protective medium.

[0019] Strain Gordonia paraffinivorans VKM Ac-3071D

[0020] Gordonia paraffinivorans strain VKM Ac-307 ID was isolated from liquid waste samples from the Surabaya wastewater treatment plant in Indonesia.

[0021] Cultural-morphological and physiological-biochemical characteristics

[0022] Colonies are orange and opaque. Cells are rod-shaped, 2-4.5 µm long. No spores were detected. Cultivation temperature was 28°C, and the incubation period was 1-2 days.

[0023] Cultivation and storage

[0024] The strain grows on media such as fish meal hydrolysate (FMH), Plate Count Agar (PCA), tributyrin agar (TBA), and mineral medium with palm oil at a final concentration of 1%. When grown on TBA, the strain forms hydrolytic zones around colonies of 1-2 mm, indicating the production of extracellular lipolytic enzymes. Growth on rich nutrient media was 1-2 days at a temperature of 4-50°C, with an optimum of 32-40°C. The strain is capable of growth on mineral medium with palm oil (1%); growth was 2-3 days at a temperature of 15-40°C, with an optimum of 32°C.

[0025] The strain Gordonia paraffmivorans VKM Ac-307 ID is maintained in laboratory conditions by periodic subcultures once every 2-3 months on agarized medium Plate Count Agar (РСА) consisting of: yeast extract 2.5 g / l; NaCl 5.0 g / l; D-glucose 1.0 g / l; dry enzymatic peptone 5.0 g / l; microbiological agar 15.0 g / l, distilled water to 1.0 l.

[0026] The strain Gordonia paraffmivorans VKM Ac-307 ID can be stored at a positive temperature of 4°C in a lyophilized state using natural skim milk as a protective medium.

[0027] Lipolytic activity of strains

[0028] The lipolytic activity of Pseudomonas protegens VKM B-3844D and Gordonia paraffmivorans VKM Ac-307 ID strains was determined by the presence of hydrolysis zones on tributryl agar (TBA). The ability to utilize palm oil was determined by culturing on a poor mineral medium supplemented with palm oil (1%). Both strains grew on TBA and formed hydrolysis zones around 1-2 mm colonies, indicating the production of extracellular lipolytic enzymes by the cells. On liquid medium with palm oil at 28°C, the maximum number of Gordonia paraffmivorans VKM Ac-3071D was 9.47xl0 8 cells / ml, maximum number of Pseudomonas protegens VKM B-3844D and 1.14x10 9 cells / ml. The temperature range for growth with palm oil was 15–40°C for Gordonia paraffmivorans VKM Ac-307 ID and 15–35°C for Pseudomonas protegens VKM B-3844D, with an optimum at 32°C and 28°C, respectively.

[0029] Each culture of the consortium of Pseudomonas protegens VKM B-3844D and Gordonia paraffinmivorans VKM Ac-3071D strains is obtained and stored separately. The strains are then combined into a consortium in a 1:1 ratio.

[0030] Lipolytic activity of a consortium of strains Pseudomonas protegens VKM B-3844D and Gordonia paraffmivorans VKM Ac-3071D

[0031] As part of the consortium, the strains Gordonia paraffmivorans VKM Ac-307 ID and Pseudomonas protegens VKM B-3844D grew actively in the presence of 5% palm oil (at least 10 8 cells / ml), the hydrolysis of the oil during cultivation was evidenced by an increase in the acid number of the oil by more than three times, compared to the control, the control values ​​of the peroxide number were 12.43 mg KOH / g, in the experimental samples during the cultivation of the consortium - 39.97 mg and 51.2 mg KOH / g.

[0032] The possibility of implementing the claimed invention is shown, but not limited, in examples of specific implementation.

[0033] Examples of preferred embodiment

[0034] Example 1. Isolation and cultivation of the Pseudomonas protegens strain VKM B-3844D

[0035] The strain was isolated from liquid waste samples from a wastewater treatment plant in Surabaya, Indonesia. A 450 ml sample of water from an aeration tank was transported to the laboratory at a positive temperature of 5±2°C. Some physicochemical parameters of the waste that served as a source for microorganism cultivation were analyzed: pH 6.9, COD 92.7 mgO / L, ammonium ion content 31.6 mg / L, nitrate ion 134 mg / L, nitrite ion 30 mg / L, sulfates 263 mg / L, chlorides 214 mg / L, phosphorus phosphates 4.16 mg / L.

[0036] The Pseudomonas protegens strain VKM B-3844D was isolated by inoculating tenfold dilutions of water onto an agarized synthetic medium containing NH4Cl - 0.625 g / l, CaCl2 - 0.0025 g / l, MnCl2 - 0.005 g / l, MgSO4x 7H2O - 0.05 g / l, FeSO4x 5H2O - 0.0025 g / l, NaCl - 1.25 g / l, Na2HPO4 - 2.5 g / l, KH2PO4 - 0.25 g / l; pH 7.5 with the addition of carboxymethylcellulose at a final concentration of 1%. Cultivation was carried out at a temperature of 28°C for 2 days. Colonies were subcultured at least 3-5 times on Plate Count Agar (PCA) medium, with culture purity monitored using microscopy. The resulting pure culture was then identified. Sequencing of the near-complete 16S rRNA gene sequence (1415 bp) and the gyrB sequence (1145 bp) was used as the primary method for species identification. Genomic DNA from the cultures was isolated using the Biolabmix (DU-50) kit according to the manufacturer's instructions (http: / / biolabmix.ru / ).To amplify bacterial 16S rRNA genes, which are universal phylogenetic markers, primers 27F [DeLong EF Archaea in coastal marine environments / / Proceedings of the National Academy of Sciences of the United States of America. - 1992. - Vol. 89, Iss. 12. - P. 5685-5689. - DOI: 10.1073 / pnas.89.12.5685] and 1492R [Weisburg WG, Bams SM, Pelletier DA, Lane DJ 16S ribosomal DNA amplification for phylogenetic study / / Journal of Bacteriology - 1991. - Vol. 173, Iss. 2. - P. 697-703] were used. - DOI: 10.1128 / jb.173.2.697-703.1991]. The 50 μl PCR mixture contained 1xPCR buffer (Biolabmix), 2.5 mM MgCl2 (Biolabmix), 0.2 mM dNTP mixture (Biolabmix), 10 pm of each primer (Synthol LLC), 0.7 units of thermostable HS-Taq polymerase (Biolabmix), 3 μl of DNA template (at a concentration exceeding 50 ng), and the final volume was made up with sterile deionized water.Amplification of the 16S rRNA genes was carried out according to the following program: initial denaturation at 95°C for 20 s; subsequent 6 cycles of denaturation at 95°C for 10 s, primer annealing at 45°C for 20 s, primer elongation at 72°C for 1.5 min; subsequent 27 cycles of denaturation at 95°C for 10 s, primer annealing at 55°C for 20 s; primer elongation at 72°C for 1.5 min; final elongation at 72°C for 3 min [Gerasimchuk A. L., Ivasenko D. A., Kasymova A. A., Frank Yu. A. Selective cultivation of bacterial strains with lipolytic and oil-oxidizing activity from the bottom sediments of the Ob River in Western Siberia / / Vavilov Journal of Genetics and Breeding. 2022. Vol. 26. No. 5. Pp. 449-457. DOI: 10.18699 / VJGB-22-55], Amplification of the gyrB genes was carried out using primers UP-1 and UP-2r [Yamamoto S., Harayama S.PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains / / Appl Environ Microbiol. 1995. V.61(3). P.1104-1109. doi: 10.1128 / aem.61.3.1104-1109.1995]. The PCR mixture was prepared as described above. Thirty amplification cycles were carried out with denaturation of template DNA at 94°C for 1 min, annealing of primers at 60°C for 1 min and elongation at 72°C for 2 min. Sequencing of the obtained DNA sequences was performed using a NANOFOR-05 genetic analyzer at the Sintol Scientific and Production Company, Moscow. The analysis of the obtained nucleotide sequences of the 16S rRNA and gyrB genes was carried out using the BIOEDIT sequence editor (http: / / www.jwbrown.mbio.ncsu.edu), the BLAST program in the NCBI GenBank database (http: / / www.ncbi.nlm.nih.gov).

[0037] The strain is capable of growth on such media as fish meal hydrolysate (FMH), Plate Count Agar (PCA), tributyrin agar (TBA), and mineral medium with palm oil at a final concentration of 1%. The strain formed hydrolysis zones on a diagnostic medium with tributyrin, indicating the production of extracellular lipolytic enzymes (lipases, esterases) by the cells. Also, indirect confirmation of lipolytic activity is positive PCR amplification of a 250 bp fragment with primers OXF1 and ACR3 [Bell PJL, Sunna A., Gibbs MD, Curach NC, Nevalainen H. and Bergquist PL Prospecting for novel lipase genes using PCR / / Microbiology - 2002. - Vol. 148. - P. 2283-2291], specific to a very diverse group of lipases of the alpha-beta hydrolase family. Growth on rich nutrient media lasted 1-2 days. The pH range for growth was 4-10, with an optimum of 8-9. The temperature range for growth was 4-35°C, with an optimum of 28-32°C.The strain is capable of growth on a mineral medium with palm oil (1%) as the only carbon source, growth takes 2-3 days, the number is 10. 8 - 10 9 cells / ml, at a temperature of 15-35°C, the optimum is 28°C, at which the maximum number is 1.14x10 9 cells / ml.

[0038] The Pseudomonas protegens VKM B-3844D strain was maintained in laboratory conditions by periodic subcultures once every 2-3 months on Plate Count Agar (PCA) medium containing the following: yeast extract - 2.5 g / l; NaCl - 5.0 g / l; D-glucose - 1.0 g / l; dry enzymatic peptone - 5.0 g / l; microbiological agar - 15.0 g / l, distilled water - up to 1.0 l.

[0039] Example 2. Method of collection (long-term) storage of the Pseudomonas protegens strain VKM B-3844D

[0040] Pseudomonas protegens strain VKM B-3844D was grown in a liquid optimal nutrient medium (e.g., PCA) by adding 5% inoculum and culturing for 24 hours at 28°C. During the exponential growth phase, the liquid culture was pelleted by centrifugation, and the supernatant was removed. Skim milk was added to the resulting pellet as a protectant. The cell suspension in the protectant was poured into glass ampoules and dehydrated by lyophilization. The vials or ampoules were stored at a positive temperature of 4°C.

[0041] Example 3. Isolation and cultivation of the Gordonia paraffinivorans strain VKM Ac-307 ID

[0042] The strain was isolated from liquid waste samples from a wastewater treatment plant in Surabaya, Indonesia. A 450-mL sample of water from an aeration tank was transported to the laboratory at a positive temperature of 5±2°C. Gordonia paraffinivorans strain VKM Ac-307 ID was isolated by inoculating tenfold dilutions of the water on agarized synthetic medium, the composition of which is described in Example 1, with the addition of carboxymethylcellulose at a final concentration of 1%. Cultivation was carried out at a temperature of 28°C for 2 days. Colonies were subcultured at least 3-5 times on solid PCA medium with culture purity control by microscopy. The resulting pure culture was then identified based on 16S rRNA gene sequence analysis, as well as on cultural, morphological, and physiological-biochemical characteristics. Sequencing of the nearly complete sequence of the 16S rRNA gene (1415 bp) was used as the main method of species identification.Genomic DNA from the cultures was isolated using the Biolabmix (DU-50) kit according to the manufacturer's instructions (htp: / / biolabmix.ru / ). For the amplification of bacterial 16S rRNA genes, which are universal phylogenetic markers, primers 27F and 1492R were used, as described for the Pseudomonas protegens strain VKM B-3844D in Example 1. Analysis of the obtained nucleotide sequences of the 16S rRNA genes was performed using the BIOEDIT sequence editor (htp: / / www.jwbrown.mbio.ncsu.edu) and the UGENE Unipro tool (htps: / / ugene.net / ru / download.html), as well as the BLAST program in the NCBI GenBank database (htp: / / www.ncbi.nlm.nih.gov).

[0043] The strain is capable of growth on media such as fish meal hydrolysate (FMH), Plate Count Agar (PCA), tributyrin agar (TBA), and mineral medium with palm oil at a final concentration of 1%. The strain formed hydrolytic zones on a diagnostic medium with tributyrin, indicating the production of extracellular lipolytic enzymes by the cells. In addition, the strain grew on a liquid synthetic medium with palm oil (1% v / v) as the sole carbon source, and at 28°C, the maximum abundance of Gordonia paraffinivorans VKM Ac-3071D was 9.47x10 8 cells / ml. On a mineral medium with palm oil (1%), growth took 2-3 days with the formation of a population of 10 8 -10 9 cells / ml at a temperature of 15–40°C, optimum 32°C.

[0044] Growth on rich nutrient media lasted 1-2 days at a temperature of 28°C. The pH range for growth was 4-10, with an optimum of 7.5. The temperature range for growth was 4-50°C, with an optimum of 32-40°C.

[0045] The strain Gordonia paraffinivorans VKM Ac-3071D was maintained in laboratory conditions by periodic subcultures once every 2-3 months on agarized medium Plate Count Agar (РСА) consisting of: yeast extract 2.5 g / l; NaCl 5.0 g / l; D-glucose 1.0 g / l; dry enzymatic peptone 5.0 g / l; microbiological agar 15.0 g / l, distilled water to 1.0 l.

[0046] Example 4. Method of collection (long-term) storage of the Gordonia paraffinivorans strain VKM Ac-307 ID

[0047] Gordonia paraffinivorans strain VKM Ac-307 ID is grown in a liquid optimal nutrient medium (e.g., PCA) by adding 5% inoculum and culturing for 24 hours at 28°C. During the exponential growth phase, the liquid culture is pelleted by centrifugation, and the supernatant is removed. Skim milk is added to the resulting pellet as a protectant. The cell suspension in the protectant is poured into glass ampoules and dehydrated by lyophilization. The vials or ampoules are stored at a positive temperature of 4°C.

[0048] Example 5. Application of a consortium of Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-307 ID for the biodegradation of palm oil

[0049] Experiments were conducted under sterile conditions in 19-liter glass flasks containing 10 liters of mineral nutrient medium supplemented with 5% palm oil. Aeration was achieved by pumping sterile air into the flasks using a compressor. A 2% inoculum (a liquid culture of Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-307 ID strains, grown separately, in a 1:1 ratio) was added to the experimental flasks. Cultivation was carried out at room temperature (22-23°C). In experiment A, to monitor the dynamics of degradation processes after 5, 10, and 15 days, visual changes in the culture fluid were recorded, residual oil was collected, and acid and peroxide values ​​were analyzed. Experiment B included an experimental flask with the consortium (experiment) and a control flask without the addition of inoculum (control), including analysis of the acid and peroxide values ​​after 15 days of incubation under the conditions described above.

[0050] According to the results of experiment A, after 5 days of cultivation, the total number of the consortium was at least 10 8 cells / ml and did not decrease until the end of experiment A. A significant portion of the palm oil was emulsified into the nutrient medium. The acid number values ​​increased compared to crude palm oil by 32, 132, and more than 240 times after 5, 10, and 15 days of the biodegradation experiment, indicating active hydrolysis of palm oil under the influence of the consortium of Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-307 ID strains.

[0051] Experiment B demonstrated that chemical hydrolysis of palm oil occurred in the control flask under aeration. However, in the experimental flask, hydrolysis was more efficient, more than 3.2 times more efficient under the influence of the consortium of strains (when comparing the acid value). Peroxide values ​​remained unchanged, suggesting that no chemical or biological formation of peroxide compounds occurred during the experiments. Furthermore, the amount of free fatty acids (C 16:1, C 18:2) increased in the culture fluid, along with changes in their composition. Under the influence of the consortium, fatty acids of the 18:0 and 18:1 types appeared, which were not detected in the chemical control without the addition of the consortium.

[0052] Example 6. Application of a consortium of Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-307 ID for the biodegradation of palm oil production waste

[0053] The storage ponds were modeled in 5-liter glass aquariums containing a mixture of 2 liters of non-sterile tap water and 100 g of raw liquid waste from a storage pond located at PT Agro Indomas, sample location coordinates -0.9141973777308665, 116.83024569376929. The raw liquid waste from the storage pond was a viscous mass with a high (89.5%) content of palm oil. Using a compressor, aeration conditions were created in two aquariums, one of which (experimental) was supplemented with a consortium of Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-307 ID strains in an amount corresponding to 10 9 cells / ml per 1 m 3liquid waste. An aquarium without aeration (control 1) and an aquarium with aeration (control 2) served as a control (without the addition of the consortium of strains). The experiment was carried out at a room temperature of about 22°C. The duration of the experiment was 33 days. Less than 24 hours later, turbidity of the water was observed in the experimental aquarium, indicating active growth of the introduced consortium and the development of bacterial biomass. Dense viscous waste with a high oil content (fat mass fraction - 89.5%), used for the experiment and analysis of its destruction, remained insoluble throughout the experiment and covered the surface of the liquid in the aquariums. However, at the end of the experiment, the appearance and consistency of the waste from the experimental aquarium with the consortium differed from the samples from the control aquariums. The results of the analysis showed that the experimental sample (experiment) contained the lowest amount of fat in dry matter (64.28%) compared to control 2 (68.5%) and control 1 (82.01%). In addition, a significant increase in free fatty acids such as C18:2 and C18:3 was revealed in the experimental aquarium (the measured peak area values ​​increased from 192.27 to 1378.97 mV sec and from 50.00 to 118.25 mV sec, respectively), indicating the destruction of fat-containing wastewater from food industry enterprises under the influence of a consortium of lipophilic bacterial strains Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-3071D. and.

[0054] Industrial applicability

[0055] Thus, a consortium of lipophilic bacterial strains, Pseudomonas protegens VKM B-3844D and Gordonia paraffinivorans VKM Ac-3071 D, was developed for the biodegradation of palm oil. Its distinguishing feature is the combination of two bacterial strains capable of active growth in a medium containing palm oil as the sole organic substance. The resulting consortium can be used in bioremediation technologies for liquid waste from palm oil production (POME).

Claims

Invention formula A consortium of lipophilic bacterial strains for the biodegradation of palm oil, consisting of Pseudomonas protegens VKM B-3844D and Gordonia parafinivorans VKM Ac-307 ID in a 1:1 ratio.