Production of controlled molecular weight hyaluronic acid by glucostat strategy using recombinant Lactococcus lactis cultures

Jeeva, Pandeeswari; Shanmuga Doss, Sreeja; Sundaram, Vijayakumar; Jayaraman, Guhan

doi:10.1007/s00253-019-09769-0

Biotechnological products and process engineering
Published: 09 April 2019

Production of controlled molecular weight hyaluronic acid by glucostat strategy using recombinant Lactococcus lactis cultures

Pandeeswari Jeeva¹^na1,
Sreeja Shanmuga Doss¹^na1,
Vijayakumar Sundaram¹ &
Guhan Jayaraman ORCID: orcid.org/0000-0003-0084-162X¹

Applied Microbiology and Biotechnology volume 103, pages 4363–4375 (2019)Cite this article

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Abstract

Hyaluronic acid (HA) is a biopolymer with wide biomedical and cosmetic applications, wherein the molecular weight of HA (MW_HA) is an important quality parameter that determines its suitability for the targeted application. To produce HA with desired molecular weight, it is important to identify parameters that offer tunability and control of MW_HA at a desired value during fermentation. In this work, two tunable parameters, viz. glucose concentration and combination of HA biosynthetic genes expressed, were used to produce HA of different molecular weights. Three recombinant strains of Lactococcus lactis were constructed, using a combination of the has-operon genes from Streptococcus zooepidemicus (hasA, hasB, hasE) and the α-phosphoglucomutase gene (pgmA) from L. lactis. Batch fermentations of these recombinant strains at different initial glucose concentrations enabled production of HA with different molecular weights. Co-expression of hasABE was observed to be particularly effective in improving the MW_HA. It was observed during batch fermentations of all these recombinant L. lactis cultures that the MW_HA decreases steadily during the later part of the fermentation and the final value is 19–43% lower than the peak MW_HA produced. Analysis of the fermentation data showed that the decrease in MW_HA correlated strongly with the decrease in specific productivity of the culture. To overcome this decrease in MW_HA, a glucostat strategy was successfully devised which could maintain a high value of specific productivity throughout the glucostat phase and result in constant-MW HA production. Glucostat processes were designed with the three recombinant L. lactis strains at two different glucose concentrations to produce constant molecular weight HA ranging from 0.4 to 1.4 MDa. This is the first report of its kind in literature that demonstrates production of controlled MW HA over a wide range by using a combination of tunable parameters and suitable process control strategies.

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Acknowledgements

The authors would like to acknowledge Indian Institute of Technology Madras for providing infrastructure to carry out this work and Ministry of Human Resource Development, Government of India for providing Fellowships to Sreeja Shanmuga Doss and Pandeeswari Jeeva.

Funding

The Department of Biotechnology (Ministry of Science and Technology, Govt. of India) provided funds to execute this project through Project No. BT/IN/Finland/30/GJ/2013.

Author information

Pandeeswari Jeeva and Sreeja Shanmuga Doss contributed equally to this work.

Authors and Affiliations

Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai, 600036, India
Pandeeswari Jeeva, Sreeja Shanmuga Doss, Vijayakumar Sundaram & Guhan Jayaraman

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Pandeeswari Jeeva
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Sreeja Shanmuga Doss
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Vijayakumar Sundaram
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Guhan Jayaraman
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Correspondence to Guhan Jayaraman.

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Jeeva, P., Shanmuga Doss, S., Sundaram, V. et al. Production of controlled molecular weight hyaluronic acid by glucostat strategy using recombinant Lactococcus lactis cultures. Appl Microbiol Biotechnol 103, 4363–4375 (2019). https://doi.org/10.1007/s00253-019-09769-0

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Received: 04 January 2019
Revised: 11 March 2019
Accepted: 12 March 2019
Published: 09 April 2019
Issue Date: 04 June 2019
DOI: https://doi.org/10.1007/s00253-019-09769-0

Keywords

Hyaluronic acid
Lactococcus lactis
Molecular weight variation
Glucostat
Constant molecular weight HA