Abstract
For strain improvement, robust and scalable high-throughput cultivation systems as well as simple and rapid high-throughput detection methods are crucial. However, most of the screening methods for lactic acid bacteria (LAB) strains were conducted in shake flasks and detected by high-performance liquid chromatography (HPLC), making the screening program laborious, time-consuming and costly. In this study, an integrated strategy for high-throughput screening of high l-lactic acid-productivity strains by Bacillus coagulans in deep-well microtiter plates (MTPs) was developed. The good agreement of fermentation results obtained in the MTPs platform with shake flasks confirmed that 24-well U-bottom MTPs could well alternate shake flasks for cell cultivation as a scale-down tool. The high-throughput pH indicator (bromocresol green) and l-lactate oxidase (LOD) assays were subsequently developed to qualitatively and quantitatively analyze l-lactic acid concentration. Together with the color halos method, the pH indicator assay and LOD assay, the newly developed three-step screening strategy has greatly accelerated the screening process for LAB strains with low cost. As a result, two high l-lactic acid-productivity mutants, IH6 and IIIB5, were successfully screened out, which presented, respectively, 42.75 and 46.10 % higher productivities than that of the parent strain in a 5-L bioreactor.
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References
Vijayakumar J, Aravindan R, Viruthagiri T (2008) Recent trends in the production, purification and application of lactic acid. Chem Biochem Eng Q 22:245–264
Abdel-Rahman MA, Tashiro Y, Sonomoto K (2013) Recent advances in lactic acid production by microbial fermentation processes. Biotechnol Adv 31:877–902
John RP, Nampoothiri KM, Pandey A (2007) Fermentative production of lactic acid from biomass: an overview on process developments and future perspectives. Appl Microbiol Biotechnol 74:524–534
Gao C, Ma CQ, Xu P (2011) Biotechnological routes based on lactic acid production from biomass. Biotechnol Adv 29:930–939
Yadav AK, Chaudhari AB, Kothari RM (2011) Bioconversion of renewable resources into lactic acid: an industrial view. Crit Rev Biotechnol 31:1–19
Bai DM, Zhao XM, Li XG, Xu SM (2004) Strain improvement of Rhizopus oryzae for over-production of l(+)-lactic acid and metabolic flux analysis of mutants. Biochem Eng J 18:41–48
Kadam SR, Patil SS, Bastawde KB, Khire JA, Gokhale DV (2006) Strain improvement of Lactobacillus delbrueckii NCIM 2365 for lactic acid production. Process Biochem 41:120–126
John RP, Nampoothiri KM (2008) Strain improvement of Lactobacillus delbrueckii using nitrous acid mutation for l-lactic acid production. World J Microbiol Biotechnol 24:3105–3109
Li SC, Zhu ZY, Gu SB, Liu HX, Wang DD (2011) Mutation-screening in l-(+)-lactic acid producing strains by ion implantation. Indian J Microbiol 51:138–143
Tsai SP, Coleman RD, Moon SH, Schneider KA, Sanville Millard C (1993) Strain screening and development for industrial lactic acid fermentation. Appl Biochem Biotechnol 39–40:323–325
Adnan AFM, Tan IKP (2007) Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential. Bioresour Technol 98:1380–1385
Playne MJ (1985) Determination of ethanol, volatile fatty acids, lactic and succinic acids in fermentation liquids by gas chromatography. J Sci Food Agric 36:638–644
Kenney BF (1991) Determination of organic acids in food samples by capillary electrophoresis. J Chromatogr A 546:423–430
Shen X, Zhang GX, Zhang DQ (2012) A new fluorometric turn-on detection of L-lactic acid based on the cascade enzymatic and chemical reactions and the abnormal fluorescent behavior of silole. Org Lett 14:1744–1747
Olson GF (1962) Optimal conditions for the enzymatic determination of l-lactic acid. Clin Chem 8:1–10
Lockridge O, Massey V, Sullivan PA (1972) Mechanism of action of the flavoenzyme lactate oxidase. J Biol Chem 247:8097–8106
Ibupoto ZH, Shah SMUA, Khun K, Willander M (2012) Electrochemical l-lactic acid sensor based on immobilized ZnO nanorods with lactate oxidase. Sensors 12:2456–2466
Liu RM, Liang LY, Ma JF, Ren XY, Jiang M, Chen KQ, Wei P, Ouyang PK (2013) An engineering Escherichia coli mutant with high succinic acid production in the defined medium obtained by the atmospheric and room temperature plasma. Process Biochem 48:1603–1609
Meng Y, Xue YF, Yu B, Gao CH, Ma YH (2012) Efficient production of l-lactic acid with high optical purity by alkaliphilic Bacillus sp. WL-S20. Bioresour Technol 116:334–339
Phrueksawan P, Kulpreecha S, Sooksai S, Thongchul N (2012) Direct fermentation of l(+)-lactic acid from cassava pulp by solid state culture of Rhizopus oryzae. Bioprocess Biosyst Eng 35:1429–1436
Li C, Xia JY, Chu J, Wang YH, Zhuang YP, Zhang SL (2013) CFD analysis of the turbulent flow in baffled shake flasks. Biochem Eng J 70:140–150
Tan J, Chu J, Hao YY, Guo YX, Zhuang YP, Zhang SL (2013) High-throughput system for screening of Cephalosporin C high-yield strain by 48-deep-well microtiter plates. Appl Biochem Biotechnol 169:1683–1695
Yu K, Hu S, Huang J, Mei LH (2011) A high-throughput colorimetric assay to measure the activity of glutamate decarboxylase. Enzyme Microb Technol 49:272–276
Banerjee A, Kaul P, Sharma R, Banerjee UC (2003) A high-throughput amenable colorimetric assay for enantioselective screening of nitrilase-producing microorganisms using pH sensitive indicators. J Biomol Screen 8:559–565
Trinder P (1969) Determination of blood glucose using an oxidase-peroxidase system with a non-carcinogenic chromogen. J Clin Pathol 22:158–161
Su F, Xu P (2014) Genomic analysis of thermophilic Bacillus coagulans strains: efficient producers for platform bio-chemicals. Sci Rep. doi:10.1038/srep03926
Vinci VA, Hoerner TD, Coffman AD, Schimmel TG, Dabora RL, Kirpekar AC, Ruby CL, Stieber RW (1991) Mutants of a lovastatin-hyperproducing Aspergillus terreus deficient in the production of sulochrin. J Ind Microbiol 8:113–119
Lin CY, Chen SH, Kou GH, Kuo CM (1999) An enzymatic microassay for lactate concentration in blood and hemolymph. Acta Zool Taiwan 10:91–101
Liaud N, Navarro D, Vidal N, Sigoillot JC, Raouche S (2014) High throughput automated colorimetric method for the screening of l-lactic acid producing microorganisms. MethodsX 1:254–257
Zhou XD, Ye LD, Wu JC (2013) Efficient production of l-lactic acid by newly isolated thermophilic Bacillus coagulans WCP10-4 with high glucose tolerance. Appl Microbiol Biotechnol 97:4309–4314
Zhang X, Zhang XF, Li HP, Wang LY, Zhang C, Xing XH, Bao CY (2014) Atmospheric and room temperature plasma (ARTP) as a new powerful mutagenesis tool. Appl Microbiol Biotechnol 98:5387–5396
Zong H, Zhan Y, Li X, Peng LJ, Feng FQ, Li D (2012) A new mutation breeding method for Streptomyces albulus by an atmospheric and room temperature plasma. Afr J Microbiol Res 6:3154–3158
Mehandjiev A, Kosturkova G, Mihov M (2001) Enrichment of Pisum sativum gene resources through combined use of physical and chemical mutagens. Israel J Plant Sci 49:279–284
Li XY, Liu RJ, Li J, Chang M, Liu YF, Jin QZ, Wang XG (2015) Enhanced arachidonic acid production from Mortierella alpina combining atmospheric and room temperature plasma (ARTP) and diethyl sulfate treatments. Bioresour Technol 177:134–140
Wang SL, Liu W, Wang HX, Lv CH (2012) Ultra high-pressure and ion implantation combined mutagenesis to improve the production of β-carotene from Red Yeast. Adv Mater Res Switz 554:1165–1169
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This work was financially supported by grants from Major State Basic Research Development Program of China (2013CB733600), the National High Technology Research and Development Program (2015AA021005) and the National Natural Science Foundation of China (Grant No. 31200024).
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Lv, X., Song, J., Yu, B. et al. High-throughput system for screening of high l-lactic acid-productivity strains in deep-well microtiter plates. Bioprocess Biosyst Eng 39, 1737–1747 (2016). https://doi.org/10.1007/s00449-016-1649-y
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DOI: https://doi.org/10.1007/s00449-016-1649-y