Biphasic Liquid-Liquid Extraction of Biosurfactant from <i>Lactobacillus delbrueckii</i>

Goyal, Surbhi; Singh, Jagdish

doi:10.1590/1678-4324-2022210325

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Article - Agriculture, Agribusiness and Biotechnology • Braz. arch. biol. technol. 65 • 2022 • https://doi.org/10.1590/1678-4324-2022210325 copy

Biphasic Liquid-Liquid Extraction of Biosurfactant from Lactobacillus delbrueckii

Authorship SCIMAGO INSTITUTIONS RANKINGS

HIGHLIGHTS

Biphasic liquid-liquid extraction of L. delbrueckii BS from the broth is executed.
Extraction parameters are optimized and partition coefficient calculated.
Co-current and countercurrent extraction of BS increased its yield.
Purified BS characterized as glycoprotein/polymeric.

Abstract:

The present work is focused mainly on optimizing the extraction of glycoprotein- biosurfactant (BS) produced by Lactobacillus delbrueckii in a biphasic liquid-liquid extraction. The production yield of BS is significantly affected by extraction strategies instead of the whole fermentation process, so it becomes a straightforward approach to enhance BS yield while optimizing various extraction parameters. The tailoring of process parameters for BS extraction was achieved by OFAT (one factor at a time) strategy and partition coefficient (K_d) served as the calculation factor for extraction. The optimal yield of BS (5+0.1 g/L) from CFB (from cell-free broth) was achieved by solvents; chloroform, methanol, and, butanol, 1:2:1 (v/v), from cell-free broth (CFB), 30% (v/v), at pH 3.5, temperature 37°C after extraction time of 60 min. Under optimized conditions, the extraction yield was 78.5% higher and subsequently, a co-current, and counter-current system enhanced the extraction yield by 16% (5.8g/L) and 20% (6.0g/L) respectively. The purity of extracted BS (EBS) was confirmed by UV/Visible spectroscopy and HPLC (High-performance liquid chromatography). The concentration-dependent activity profile of EBS analyzed by ODA (oil displacement area; 50.24+0.3 cm²), DCD (drop collapse diameter; 1.1+0.3 cm), and EI₂₄ (73+0.3%), exhibited enhancement by 60, 52, and 55% respectively as compared to control. Thin-layer chromatography (TLC), FTIR (Fourier Transform Infra-Red) and NMR (Nuclear Magnetic Resonance) techniques confirmed the polymeric glycoprotein (65:35 protein: carbohydrates %) nature of BS.

Keywords:
Polymeric biosurfactant; Lactobacillus delbrueckii; Liquid-liquid extraction; biphasic; co-current/countercurrent.

Stages	Co-current		Counter current
	K_d	Extraction yield (g/L)	K_d	Extraction yield (g/L)
I	7.2+0.1	5.0+0.2	6.9+0.2	3.5+0.4
II	1.8+0.3	0.5+0.1	7.6+0.1	4.8+0.2
III	1.5+0.1	0.2+0.1	7.8+0.2	5.9+0.4
Overall	7.1+0.2	5.8+0.2	7.4+0.2	6.0+0.3

Assignments	H¹- NMR (ppm) Peak values	Reference
CH₂=CH	7.35	[²⁹29. Anjum F, Gautam G, Edgard G, Negi S. Biosurfactant production through Bacillus sp. MTCC 5877 and its multifarious applications in food industry. Biores Technol. 2016;213:262-9.]
Proton attached to C-1 of sugar moiety	4.05	[⁴⁵45. Jain RM, Mody K, Joshi N, Mishra A, Jha B. Production and structural characterization of biosurfactant produced by an alkaliphilic bacterium, Klebsiella sp.: Evaluation of different carbon sources. Colloids Surfaces B Biointerfaces. 2013;108:199-204. https://doi.org/10.1016/j.colsurfb.2013.03.002 https://doi.org/10.1016/j.colsurfb.2013.... ]
Proton attached to C-2 of sugar moiety	3.50	[⁴⁶46. Miao M, Ma Y, Huang C, Jiang B, Cui SW, Zhang T. Physicochemical properties of a water soluble extracellular homopolysaccharide from Lactobacillus reuteri SK24.003. Carbohydr Polym. 2015;131:377-83. https://doi.org/10.1016/J.CARBPOL.2015.05.066 https://doi.org/10.1016/J.CARBPOL.2015.0... ]
Proton attached to C-3 of sugar moiety	3.50	[⁴⁰40. Bakhshi N, Sheikh-Zeinoddin M, Soleimanian-Zad S. Production and Partial Characterization of a Glycoprotein Bioemulsifier Produced by Lactobacillus plantarum subsp. plantarum PTCC 1896. J Agri Sci Technol. 2018;20:37-49.]
Proton attached to C-4 of sugar moiety	2.37	[²⁹29. Anjum F, Gautam G, Edgard G, Negi S. Biosurfactant production through Bacillus sp. MTCC 5877 and its multifarious applications in food industry. Biores Technol. 2016;213:262-9.]
Protein group of glucan-protein structure	1.03,1.05,1.36	[⁴⁷47. Sharma D, Ansari MJ, Gupta S, Ghamdi A Al, Pruthi P, Pruthi V. Structural characterization and antimicrobial activity of a biosurfactant obtained from bacillus pumilus DSVP18 grown on potato peels. Jundishapur J Microbiol. 2015;8. https://doi.org/10.5812/jjm.21257 https://doi.org/10.5812/jjm.21257... ]; [²⁵25. Dikit P, Methacanon P, Visessanguan W, Aran H, Maneerat S. Characterization of an unexpected bioemulsifier from spent yeast obtained from Thai traditional liquor distillation. Int J Biol Macromol. 2010;47:465-70.]

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