Acid adaptation of Lactobacillus helveticus during continuous cultures to improve undissociated lactic acid production

Lauranne Collet , Jérôme Delettre , Violaine Athès , Caroline Pénicaud , Catherine Béal

Bioresources and Bioprocessing ›› 2026, Vol. 13 ›› Issue (1) : 30

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Bioresources and Bioprocessing ›› 2026, Vol. 13 ›› Issue (1) :30 DOI: 10.1186/s40643-026-01019-2
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Acid adaptation of Lactobacillus helveticus during continuous cultures to improve undissociated lactic acid production
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Abstract

This study aims at improving the undissociated lactic acid production by Lactobacillus helveticus using a whey-based fermentation. It first describes the effect of pH on the ability of this bacterium to produce lactic acid, by considering final lactic acid concentration, production rate, volumetric productivity and sugar consumption. As a low performance was achieved at pH 4.3, an adaptive evolution of Lb. helveticus LH-B01 to acidic conditions was performed during continuous cultures of sweet hydrolysed whey. Two mutants have been isolated, which exhibited different characteristics. The mutant Lb. helveticus LH-B01-B4 displayed the higher maximal total lactic acid concentration (37.9 g/L), sugar consumption (82%) and volumetric productivity (0.39 g/L/h), when compared to the parental strain and the mutant Lb. helveticus LH-B01-A4. This performance was explained by the higher critical undissociated lactic acid concentration (10.1 g/L) of Lb. helveticus LH-B01-B4, compared with those of the parental strain (8.7 g/L) and the mutant Lb. helveticus LH-B01-A4 (7.5 g/L). From these results, the mutant strain Lb. helveticus LH-B01-B4 was the most promising option to produce undissociated lactic acid during low pH fermentation, thus making it suitable for industrial use as a descaling agent and biocide in detergents.

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Adaptive evolution / Continuous cultures / Critical undissociated lactic acid concentration / Dilution rate / Lactic acid bacteria / Acidic pH / Whey

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Lauranne Collet, Jérôme Delettre, Violaine Athès, Caroline Pénicaud, Catherine Béal. Acid adaptation of Lactobacillus helveticus during continuous cultures to improve undissociated lactic acid production. Bioresources and Bioprocessing, 2026, 13(1): 30 DOI:10.1186/s40643-026-01019-2

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References

[1]

Abdel-Rahman MA, Tashiro Y, Sonomoto K. Recent advances in lactic acid production by microbial fermentation processes. Biotechnol Adv. 2013, 31(6): 877-902.

[2]

Aeschlimann A, Von Stockar U. The production of lactic acid from whey permeate by Lactobacillus helveticus. Biotechnol Lett. 1989, 11(3): 195-200.

[3]

Alves De Oliveira R, Komesu A, Vaz Rossell CE, Maciel Filho R. Challenges and opportunities in lactic acid bioprocess design—from economic to production aspects. Biochem Eng J. 2018, 133: 219-239.

[4]

Amrane A. Batch cultures of supplemented whey permeate using Lactobacillus helveticus : unstructured model for biomass formation, substrate consumption and lactic acid production. Enzyme Microb Technol. 2001, 28(9–10): 827-834.

[5]

Amrane A, Prigent Y. Influence of media composition on lactic acid production rate from whey by Lactobacillus helveticus. Biotechnol Lett. 1993, 15(3): 239-244.

[6]

Amrane A, Prigent Y. Influence of an initial addition of lactic acid on growth, acid production and their coupling for batch cultures of Lactobacillus helveticus. Bioprocess Eng. 1998, 19(4): 307-312.

[7]

Béal C, Corrieu G. On-line indirect measurements of biological variables and their kinetics during pH controlled batch cultures of thermophilic lactic acid bacteria. J Food Eng. 1995, 26(4): 511-525.

[8]

Béal C, Arana-Agudelo P, Farel T, Moussa M, Athès V (2023) 5 - Lactic acid microbial production and recovery: review and recent advances in bioprocess integration. In: Montet D, Ray RC, De Carvalho Azevedo VA, Paramithiotis S (eds) Lactic acid bacteria as cell factories. Woodhead Publishing, pp 77–108

[9]

Castillo Martinez FA, Balciunas EM, Salgado JM, Domínguez González JM, Converti A, Oliveira RPDS. Lactic acid properties, applications and production: a review. Trends Food Sci Technol. 2013, 30(1): 70-83.

[10]

Fitzpatrick JJ, O’Keeffe U. Influence of whey protein hydrolysate addition to whey permeate batch fermentations for producing lactic acid. Process Biochem. 2001, 37(2): 183-186.

[11]

Gätje G, Gottschalk G. Limitation of growth and lactic acid production in batch and continuous cultures of Lactobacillus helveticus. Appl Microbiol Biotechnol. 1991, 34(4): 446-449.

[12]

Ghosh BC, Prasad LN, Saha NP. Enzymatic hydrolysis of whey and its analysis. J Food Sci Technol. 2017, 54(6): 1476-1483.

[13]

Gresham D, Dunham MJ. The enduring utility of continuous culturing in experimental evolution. Genomics. 2014, 104(6): 399-405.

[14]

Griffiths MW, Tellez AM. Lactobacillus helveticus: the proteolytic system. Front Microbiol. 2013.

[15]

Hartono S, Meijerink MFA, Abee T, Smid EJ, Van Mastrigt O. The stressostat: a novel approach in adaptive laboratory evolution to improve end-product resistance. New Biotechnol. 2023, 78: 123-130.

[16]

Hill AR, Irvine DM, Bullock DH. Buffer capacity of cheese wheys. J Food Sci. 1985, 50(3): 733-738.

[17]

Huang J, Huang L, Lin J, Xu Z, Cen P. Organic chemicals from bioprocesses in China. Adv Biochem Eng Biotechnol. 2010, 122: 43-71.

[18]

Kim J, Kim Y-M, Lebaka VR, Wee Y-J. Lactic acid for green chemical industry: recent advances in and future prospects for production technology, recovery, and applications. Fermentation. 2022, 8(11. 609

[19]

Komesu A, Oliveira JARde, Martin LHdaS, Wolf Maciel MR, Maciel Filho R. Lactic acid production to purification : a review. BioResources. 2017, 122): 4364-4383.

[20]

Konings WN. Siezen RJ, Kok J, Abee T, Schasfsma G. The cell membrane and the struggle for life of lactic acid bacteria. Lactic acid bacteria: genetics, metabolism and applications. 2002, Berlin, Springer327.

[21]

Li Z, Zhang L, Zhang B, Bao J. pH shifting adaptive evolution stimulates the low pH tolerance of Pediococcus acidilactici and high L-lactic acid fermentation efficiency. Bioresour Technol. 2025, 416. 131813

[22]

Mavrommati M, Daskalaki A, Papanikolaou S, Aggelis G. Adaptive laboratory evolution principles and applications in industrial biotechnology. Biotechnol Adv. 2022, 54. 107795

[23]

Paillet T, Lossouarn J, Figueroa C, Midoux C, Rué O, Petit M-A, Dugat-Bony E. Virulent phages isolated from a smear-ripened cheese are also detected in reservoirs of the cheese factory. Viruses. 2022, 14(8. 1620

[24]

Roy D, Goulet J, LeDuy A. Batch fermentation of whey ultrafiltrate by Lactobacillus helveticus for lactic acid production. Appl Microbiol Biotechnol. 1986, 243): 206-213.

[25]

Sauer U (2001) Evolutionary engineering of industrially important microbial phenotypes. In: Metabolic engineering. Springer. https://doi.org/10.1007/3-540-45300-8_7

[26]

Schepers AW, Thibault J, Lacroix C. Lactobacillus helveticus growth and lactic acid production during pH-controlled batch cultures in whey permeate/yeast extract medium. Part II: kinetic modeling and model validation. Enzyme Microb Technol. 2002, 30(2): 187-194.

[27]

Singhvi M, Zendo T, Sonomoto K. Free lactic acid production under acidic conditions by lactic acid bacteria strains : challenges and future prospects. Appl Microbiol Biotechnol. 2018, 102(14): 5911-5924.

[28]

Tsermoula P, Khakimov B, Nielsen JH, Engelsen SB. Whey—the waste-stream that became more valuable than the food product. Trends Food Sci Technol. 2021, 118: 230-241.

[29]

Wolfschoon Pombo AF, Spiegel TL, Hernandez-Zenil E. Buffering curves of ideal whey fractions obtained from a cascade membrane separation process. Int J Dairy Technol. 2017, 70(2): 287-296.

[30]

Zhang J, Wu C, Du G, Chen J. Enhanced acid tolerance in Lactobacillus casei by adaptive evolution and compared stress response during acid stress. Biotechnol Bioprocess Eng. 2012, 17(2): 283-289.

[31]

Zhang W, Tao Y, Wu M, Xin F, Dong W, Zhou J, Gu J, Ma J, Jiang M. Adaptive evolution improves acid tolerance and succinic acid production in Actinobacillus succinogenes. Process Biochem. 2020, 98: 76-82.

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