Low cost and sustainable hyaluronic acid production in a manufacturing platform based on Bacillus subtilis 3NA strain.
Bacillus subtilis
Fermentation
Green chemistry
Hyaluronic acid
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
Apr 2021
Apr 2021
Historique:
received:
02
02
2021
accepted:
16
03
2021
revised:
09
03
2021
pubmed:
6
4
2021
medline:
15
5
2021
entrez:
5
4
2021
Statut:
ppublish
Résumé
Hyaluronic acid (HA) is a high value glycosaminoglycan mostly used in health and cosmetic applications. Commercial HA is produced from animal tissues or in toxigenic bacteria of the genus Streptococcus grown in complex media, which are expensive and raise environmental concerns due to the disposal of large amounts of broth with high organic loads. Other microorganisms were proposed as hosts for the heterologous production of HA, but the methods are still costly. The extraordinary capacity of this biopolymer to bind and retain water attracts interest for large-scale applications where biodegradable materials are needed, but its high cost and safety concerns are barriers for its adoption. Bacillus subtilis 3NA strain is prototrophic, amenable for genetic manipulation, GRAS, and can rapidly reach high cell densities in salt-based media. These phenotypic traits were exploited to create a platform for biomolecule production using HA as a proof of concept. First, the 3NA strain was engineered to produce HA; second, a chemically defined medium was formulated using commodity-priced inorganic salts combined at the stoichiometric ratios needed to build the necessary quantities of biomass and HA; and third, a scalable fermentation process, where HA can be produced at the maximum volumetric productivity (VP), was designed. A comparative economic analysis against other methods indicates that the new process may increase the operating profit of a manufacturing plant by more than 100%. The host, the culture medium, and the rationale employed to develop the fermentation process described here, introduce an IP-free platform that could be adaptable for production of other biomolecules. KEY POINTS: • A biomolecule production platform based on B. subtilis 3NA strain and a synthetic medium was tested for hyaluronic acid biosynthesis • A fermentation process with the maximum volumetric productivity was designed • A techno-economic analysis forecasts a significant reduction in the manufacturing cost compared to the current methods.
Identifiants
pubmed: 33818671
doi: 10.1007/s00253-021-11246-6
pii: 10.1007/s00253-021-11246-6
doi:
Substances chimiques
Culture Media
0
Hyaluronic Acid
9004-61-9
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
3075-3086Subventions
Organisme : Glyco@Alps
ID : ANR-15-IDEX-02
Commentaires et corrections
Type : ErratumIn
Références
Anagnostopoulos C, Spizizen J (1961) Requirements for transformation in Bacillus subtilis. J Bacteriol 81(5):741–746. https://doi.org/10.1128/JB.81.5.741-746.1961
doi: 10.1128/JB.81.5.741-746.1961
pubmed: 16561900
pmcid: 279084
Ben-Arye T, Levenberg S (2019) Tissue engineering for clean meat production. Front Sustain Food Syst 3(46). https://doi.org/10.3389/fsufs.2019.00046
Blumenkrantz N, Asboe-Hansen G (1973) New method for quantitative determination of uronic acids. Anal Biochem 54(2):484–489. https://doi.org/10.1016/0003-2697(73)90377-1
doi: 10.1016/0003-2697(73)90377-1
pubmed: 4269305
Boeriu CG, Springer J, Kooy FK, van den Broek LAM, Eggink G (2013) Production methods for hyaluronan. Int J Carbohydr Chem 2013:624967–624914. https://doi.org/10.1155/2013/624967
doi: 10.1155/2013/624967
Chen X, Zhang C, Cheng J, Shi X, Li L, Zhang Z, Bai J, Chen Y, Li S, Ying H (2013) Enhancement of adenosine production by Bacillus subtilis CGMCC 4484 through metabolic flux analysis and simplified feeding strategies. Bioproc Biosyst Eng 36(12):1851–1859. https://doi.org/10.1007/s00449-013-0959-6
doi: 10.1007/s00449-013-0959-6
Chien LJ, Lee CK (2007a) Enhanced hyaluronic acid production in Bacillus subtilis by coexpressing bacterial hemoglobin. Biotechnol Prog 23(5):1017–1022. https://doi.org/10.1021/bp070036w
doi: 10.1021/bp070036w
pubmed: 17691809
Chien LJ, Lee CK (2007b) Hyaluronic acid production by recombinant Lactococcus lactis. Appl Microbiol Biotechnol 77(2):339–346. https://doi.org/10.1007/s00253-007-1153-z
doi: 10.1007/s00253-007-1153-z
pubmed: 17805528
Dahod SK, Greasham R, Kennedy M (2010) Raw materials selection and medium development for industrial fermentation processes. In: Bull AT, Junker B, Katz L, Lynd LR, Masurekar P, Reeves CD, Zhao H (eds) Manual of Industrial Microbiology and Biotechnology 3rd edn. Am Soc Microbiol
Dauner M, Storni T, Sauer U (2001) Bacillus subtilis metabolism and energetics in carbon-limited and excess-carbon chemostat culture. J Bacteriol 183(24):7308–7317. https://doi.org/10.1128/jb.183.24.7308-7317.2001
doi: 10.1128/jb.183.24.7308-7317.2001
pubmed: 11717290
pmcid: 95580
de Oliveira JD, Carvalho LS, Gomes AM, Queiroz LR, Magalhães BS, Parachin NS (2016) Genetic basis for hyper production of hyaluronic acid in natural and engineered microorganisms. Microb Cell Factories 15(1):119. https://doi.org/10.1186/s12934-016-0517-4
doi: 10.1186/s12934-016-0517-4
de Souza AB, Chaud MV, Santana MHA (2019) Hyaluronic acid behavior in oral administration and perspectives for nanotechnology-based formulations: a review. Carbohydr Polym 222:115001. https://doi.org/10.1016/j.carbpol.2019.115001
doi: 10.1016/j.carbpol.2019.115001
pubmed: 31320101
DeAngelis PL, Papaconstantinou J, Weigel PH (1993) Molecular cloning, identification, and sequence of the hyaluronan synthase gene from group A Streptococcus pyogenes. J Biol Chem 268(26):19181–19184
doi: 10.1016/S0021-9258(19)36494-4
Dicker KT, Gurski LA, Pradhan-Bhatt S, Witt RL, Farach-Carson MC, Jia X (2014) Hyaluronan: a simple polysaccharide with diverse biological functions. Acta Biomater 10(4):1558–1570. https://doi.org/10.1016/j.actbio.2013.12.019
doi: 10.1016/j.actbio.2013.12.019
pubmed: 24361428
Don MM, Shoparwe NF (2010) Kinetics of hyaluronic acid production by Streptococcus zooepidemicus considering the effect of glucose. Biochem Eng J 49(1):95–103. https://doi.org/10.1016/j.bej.2009.12.001
doi: 10.1016/j.bej.2009.12.001
Fallacara A, Baldini E, Manfredini S, Vertuani S (2018) Hyaluronic acid in the third millennium. Polymers 10(7). https://doi.org/10.3390/polym10070701
Ferreira RG, Azzoni AR, Freitas S (2018) Techno-economic analysis of the industrial production of a low-cost enzyme using E. coli: the case of recombinant β-glucosidase. Biotechnol Biofuels 11(1):81. https://doi.org/10.1186/s13068-018-1077-0
doi: 10.1186/s13068-018-1077-0
pubmed: 29610578
pmcid: 5875018
Hoffmann J, Altenbuchner J (2014) Hyaluronic acid production with Corynebacterium glutamicum: effect of media composition on yield and molecular weight. J Appl Microbiol 117(3):663–678. https://doi.org/10.1111/jam.12553
doi: 10.1111/jam.12553
pubmed: 24863652
Huang H, Ridgway D, Gu T, Moo-Young M (2003) A segregated model for heterologous amylase production by Bacillus subtilis. Enzym Microb Technol 32(3):407–413. https://doi.org/10.1016/S0141-0229(02)00312-5
doi: 10.1016/S0141-0229(02)00312-5
Huang H, Ridgway D, Gu T, Moo-Young M (2004) Enhanced amylase production by Bacillus subtilis using a dual exponential feeding strategy. Bioproc Biosyst Eng 27(1):63–69. https://doi.org/10.1007/s00449-004-0391-z
doi: 10.1007/s00449-004-0391-z
Jia Y, Zhu J, Chen X, Tang D, Su D, Yao W, Gao X (2013) Metabolic engineering of Bacillus subtilis for the efficient biosynthesis of uniform hyaluronic acid with controlled molecular weights. Bioresour Technol 132:427–431. https://doi.org/10.1016/j.biortech.2012.12.150
doi: 10.1016/j.biortech.2012.12.150
pubmed: 23433979
Jin P, Kang Z, Yuan P, Du G, Chen J (2016) Production of specific-molecular-weight hyaluronan by metabolically engineered Bacillus subtilis 168. Metab Eng 35:21–30. https://doi.org/10.1016/j.ymben.2016.01.008
doi: 10.1016/j.ymben.2016.01.008
pubmed: 26851304
Kim J-H, Yoo S-J, Oh D-K, Kweon Y-G, Park D-W, Lee C-H, Gil G-H (1996) Selection of a Streptococcus equi mutant and optimization of culture conditions for the production of high molecular weight hyaluronic acid. Enzym Microb Technol 19(6):440–445. https://doi.org/10.1016/S0141-0229(96)00019-1
doi: 10.1016/S0141-0229(96)00019-1
Kumari K, Weigel PH (1997) Molecular cloning, expression, and characterization of the authentic hyaluronan synthase from group C Streptococcus equisimilis. J Biol Chem 272(51):32539–32546. https://doi.org/10.1074/jbc.272.51.32539
doi: 10.1074/jbc.272.51.32539
pubmed: 9405467
Kurane R, Nohata Y (1994) A new water-absorbing polysaccharide from Alcaligenes latus. Biosci Biotechnol Biochem 58(2):235–238. https://doi.org/10.1271/bbb.58.235
doi: 10.1271/bbb.58.235
Lee SY (1996) High cell-density culture of Escherichia coli. Trends Biotechnol 14(3):98–105. https://doi.org/10.1016/0167-7799(96)80930-9
doi: 10.1016/0167-7799(96)80930-9
pubmed: 8867291
Li Y, Shi Z, Shao Y, Wu M, Li G, Ma T (2020) Temperature-controlled molecular weight of hyaluronic acid produced by engineered Bacillus subtilis. Biotechnol Lett 43:271–277. https://doi.org/10.1007/s10529-020-03001-0
doi: 10.1007/s10529-020-03001-0
pubmed: 32910358
Looser V, Lüthy D, Straumann M, Hecht K, Melzoch K, Kovar K (2017) Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P. pastoris: functional characterisation of a novel promoter. Appl Microbiol Biotechnol 101(8):3163–3176. https://doi.org/10.1007/s00253-017-8123-x
doi: 10.1007/s00253-017-8123-x
pubmed: 28130631
pmcid: 5380701
Lou J, Stowers R, Nam S, Xia Y, Chaudhuri O (2018) Stress relaxing hyaluronic acid-collagen hydrogels promote cell spreading, fiber remodeling, and focal adhesion formation in 3D cell culture. Biomaterials 154:213–222. https://doi.org/10.1016/j.biomaterials.2017.11.004
doi: 10.1016/j.biomaterials.2017.11.004
pubmed: 29132046
Martínez A, Ramírez OT, Valle F (1998) Effect of growth rate on the production of β-galactosidase from Escherichia coli in Bacillus subtilis using glucose-limited exponentially fedbatch cultures. Enzym Microb Technol 22(6):520–526. https://doi.org/10.1016/S0141-0229(97)00248-2
doi: 10.1016/S0141-0229(97)00248-2
Oe M, Sakai S, Yoshida H, Okado N, Kaneda H, Masuda Y, Urushibata O (2017) Oral hyaluronan relieves wrinkles: a double-blinded, placebo-controlled study over a 12-week period. Clin Cosmet Investig Dermatol 10:267–273. https://doi.org/10.2147/ccid.S141845
doi: 10.2147/ccid.S141845
pubmed: 28761365
pmcid: 5522662
Pierce JA, Robertson CR, Leighton TJ (1992) Physiological and genetic strategies for enhanced subtilisin production by Bacillus subtilis. Biotechnol Prog 8(3):211–218. https://doi.org/10.1021/bp00015a006
doi: 10.1021/bp00015a006
pubmed: 1368258
Reisman HB (2019) Economic analysis of fermentation processes. CRC Press
Reuß DR, Schuldes J, Daniel R, Altenbuchner J (2015) Complete genome sequence of Bacillus subtilis subsp. subtilis strain 3NA. Genome Announc 3(2):e00084–e00015. https://doi.org/10.1128/genomeA.00084-15
doi: 10.1128/genomeA.00084-15
pubmed: 25767229
pmcid: 4357751
Rocha EPC, Danchin A, Viari A (1999) Translation in Bacillus subtilis: roles and trends of initiation and termination, insights from a genome analysis. Nucleic Acids Res 27(17):3567–3576. https://doi.org/10.1093/nar/27.17.3567
doi: 10.1093/nar/27.17.3567
pubmed: 10446248
pmcid: 148602
Sakthiselvan P, Meenambiga SS, Madhumathi R (2019) Kinetic studies on cell growth. In: Vikas B (ed) Cell Growth. IntechOpen
Stanbury PF, Whitaker A, Hall SJ (2017) Chapter 4 - media for industrial fermentations. In: Stanbury PF, Whitaker A, Hall SJ (eds) Principles of fermentation technology, 3rd edn. Butterworth-Heinemann, Oxford, pp 213–272
doi: 10.1016/B978-0-08-099953-1.00004-1
Weissmann B, Meyer K (1954) The structure of hyalobiuronic acid and of hyaluronic acid from umbilical cord 1,2. J Am Chem Soc 76(7):1753–1757. https://doi.org/10.1021/ja01636a010
doi: 10.1021/ja01636a010
Westbrook AW, Ren X, Oh J, Moo-Young M, Chou CP (2018) Metabolic engineering to enhance heterologous production of hyaluronic acid in Bacillus subtilis. Metab Eng 47:401–413. https://doi.org/10.1016/j.ymben.2018.04.016
doi: 10.1016/j.ymben.2018.04.016
pubmed: 29698777
Widner B, Behr R, Von Dollen S, Tang M, Heu T, Sloma A, Sternberg D, Deangelis PL, Weigel PH, Brown S (2005) Hyaluronic acid production in Bacillus subtilis. Appl Environ Microbiol 71(7):3747–3752. https://doi.org/10.1128/aem.71.7.3747-3752.2005
doi: 10.1128/aem.71.7.3747-3752.2005
pubmed: 16000785
pmcid: 1168996
Wu F-C, Chou S-Z, Shih I-L (2013) Factors affecting the production and molecular weight of levan of Bacillus subtilis natto in batch and fed-batch culture in fermenter. J Taiwan Inst Chem Eng 44(6):846–853. https://doi.org/10.1016/j.jtice.2013.03.009
doi: 10.1016/j.jtice.2013.03.009
Yao J, Xu H, Shi N, Cao X, Feng X, Li S, Ouyang P (2010) Analysis of carbon metabolism and improvement of γ-polyglutamic acid production from Bacillus subtilis NX-2. Appl Biochem Biotechnol 160(8):2332–2341. https://doi.org/10.1007/s12010-009-8798-2
doi: 10.1007/s12010-009-8798-2
pubmed: 19866376
Zając M, Kulawik P, Tkaczewska J, Migdał W, Filipczak-Fiutak M, Fiutak G (2017) The effect of hyaluronic acid addition on the properties of smoked homogenised sausages. J Sci Food Agric 97(8):2316–2326. https://doi.org/10.1002/jsfa.8041
doi: 10.1002/jsfa.8041
pubmed: 27633533
Zhang L, Huang H, Wang H, Chen J, Du G, Kang Z (2016) Rapid evolution of hyaluronan synthase to improve hyaluronan production and molecular mass in Bacillus subtilis. Biotechnol Lett 38(12):2103–2108. https://doi.org/10.1007/s10529-016-2193-1
doi: 10.1007/s10529-016-2193-1
pubmed: 27565668