Skip to main content
SpringerLink
Log in

Improvement of tacrolimus production in Streptomyces tsukubaensis by mutagenesis and optimization of fermentation medium using Plackett–Burman design combined with response surface methodology

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Objective

This study was conducted to enhance the production of tacrolimus in Streptomyces tsukubaensis by strain mutagenesis and optimization of the fermentation medium.

Results

A high tacrolimus producing strain S. tsukubaensis FIM-16-06 was obtained by ultraviolet mutagenesis coupled with atmospheric and room temperature plasma mutagenesis.Then, nine variables were screened using Plackett–Burman experimental design, in which soluble starch, peptone and Tween 80 showed significantly affected tacrolimus production. Further studies were carried out employing central composite design to elucidate the mutual interaction between the variables and to work out optimal fermentation medium composition for tacrolimus production. The optimum fermentation medium was found to contain 61.61 g/L of soluble starch, 20.61 g/L of peptone and 30.79 g/L of Tween 80. In the optimized medium, the production of tacrolimus reached 1293 mg/L in shake-flask culture, and reached 1522 mg/L while the scaled-up fermentation was conducted in a 1000 L fermenter, which was about 3.7 times higher than that in the original medium.

Conclusions

Combining compound mutation with rational medium optimization is an effective approach for improving tacrolimus production, and the optimized fermentation medium could be efficiently used for industrial production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig.5
Fig. 6

Similar content being viewed by others

References

  • Allison AC (2000) Immunosuppressive drugs: the first 50 years and a glance forward. Immunopharmacology 47:63–83

    Article  CAS  Google Scholar 

  • Barreiro C, Martinez-Castro M (2014) Trends in the biosynthesis and production of the immunosuppressant tacrolimus (FK506). Appl Microbiol Biotechnol 2:497–507

    Article  Google Scholar 

  • Bhagwat PK, Jhample SB, Dandge PB (2015) Statistical medium optimization for production of collagenolytic protease by Pseudomonas sp. SUK using response surface methodology. Microbiol 84:520–530

    Article  CAS  Google Scholar 

  • Chaudhary P, Chhokar V, Choudhary P, Kumar A, Beniwal V (2017) Optimization of chromium and tannic acid bioremediation by Aspergillus niveus using Plackett–Burman design and response surface methodology. AMB Express 1:201

    Article  Google Scholar 

  • Cheng W, Di H, Liang S (2018) Identification and metabolomic analysis of chemical elicitors for tacrolimus accumulation in Streptomyces tsukubaensis. Appl Microbiol Biot 17:7541–7553

    Google Scholar 

  • Ekpenyong MG, Antai SP, Asitok AD, Ekpo BO (2017) Plackett–Burman design and response surface optimization of medium trace nutrients for glycolipopeptide biosurfactant production. Iran Biomed J 4:249–260

    Google Scholar 

  • Ekpenyong M, Asitok A, Antigha R, Ogarekpe N, Antai S (2021) Bioprocess optimization of nutritional parameters for enhanced anti-leukemic L-asparaginase production by Aspergillus candidus UCCM 00117: a sequential statistical approach. Int J Pept Res Ther 27:1501–1527

    Article  CAS  Google Scholar 

  • Elsayed EA, Farid MA, El-Enshasy HA (2019) Enhanced natamycin production by Streptomyces natalensis in shake-flasks and stirred tank bioreactor under batch and fed-batch conditions. BMC Biotechnol 19(1):46

    Article  Google Scholar 

  • Goto T, Kino T, Hatanaka H, Nishiyama M, Okuhara M, Kohsaka M, Kohsaka M, Aoki H, Imanaka H (1987) Discovery of FK-506, a novel immunosuppressant isolated from Streptomyces tsukubaensis. Transpl Proceed 5:4–8

    Google Scholar 

  • Huang D, Xia M, Li S, Wen J, Jia X (2013) Enhancement of FK-506 production by engineering secondary pathways of Streptomyces tsukubaensis and exogenous feeding strategies. J Ind Microbiol Biotechnol 9:1023–1037

    Article  Google Scholar 

  • Huang J, Ou Y, Zhang D, Zhang G, Pan Y (2018) Optimization of the culture condition of Bacillus mucilaginous using Agaricus bisporus industrial wastewater by Plackett–Burman combined with box-Behnken response surface method. AMB Express 8:141

    Article  Google Scholar 

  • Husain S, Singh N (2002) The impact of novel immunosuppressive agents on infections in organ transplant recipients and the interactions of these agents with antimicrobials. Clin Infect Dis 35:53–61

    Article  CAS  Google Scholar 

  • Jiang H, Wang Y, Guo Y, Shen J, Zhang X, Luo H, Ren N, Jiang X, Li Y (2015) An acyltransferase domain of FK506 polyketide synthase recognizing both an acyl carrier protein and coenzyme A as acyl donors to transfer allylmalonyl and ethylmalonyl units. FEBS J 13:2527–2539

    Article  Google Scholar 

  • Joyce AP, Leung SS (2013) Use of response surface methods and path of steepest ascent to optimize ligand-binding assay sensitivity. J Immunol Methods 1–2:12–23

    Article  Google Scholar 

  • Jung S, Moon S, Lee K, Park Y, Yoon S, Yoo Y (2009) Strain development of Streptomyces sp. for tacrolimus production using sequential adaptation. J Ind Microbiol Biotechnol 12:1467–1471

    Article  Google Scholar 

  • Kim HS, Park YI (2008) Isolation and identification of a novel microorganism producing the immunosuppressant tacrolimus. J Biosci Bioeng 4:418–421

    Article  Google Scholar 

  • Kol S, Merlo ME, Scheltema RA, de Vries M, Vonk RJ, Kikkert NA, Dijkhuizen L, Breitling R, Takano E (2010) Metabolomic characterization of the salt stress response in Streptomyces coelicolor. Appl Environ Microbiol 76:2574

    Article  CAS  Google Scholar 

  • Kuo CH, Hsiao FW, Chen JH, Hsieh CW, Liu YC, Shieh CJ (2013) Kinetic aspects of ultrasound-accelerated lipase catalyzed acetylation and optimal synthesis of 4’-acetoxyresveratrol. Ultrason Sonochem 20:546–552

    Article  CAS  Google Scholar 

  • Martínez-Castro M, Salehi-Najafabadi Z, Romero F, Pérez-Sanchiz R, Fernández-Chimeno RI, Martín JF, Barreiro C (2013) Taxonomy and chemically semi-defined media for the analysis of the tacrolimus producer ‘Streptomyces tsukubaensis’. Appl Microbiol Biotechnol 97(5):2139–52152. https://doi.org/10.1007/s00253-012-4364-x

    Article  CAS  PubMed  Google Scholar 

  • Mishra A, Verma S (2012) Optimization of process parameters for tacrolimus (FK 506) production by new isolate of Streptomyces sp. using response surface methodology. J Biochem Technol 3:419–425

    CAS  Google Scholar 

  • Mo SJ, Nan Y-H, Park W-J, Yoo YJ, Yoon YJ (2009) Enhanced FK506 production in Streptomyces clavuligerus CKD1119 by engineering the supply of methylmalonyl-CoA precursor. J Ind Microbiol Biotechnol 36:1473–1482

    Article  CAS  Google Scholar 

  • Mo S, Lee S-K, Jin YY, Oh CH, Suh JW (2013) Application of a combined approach involving classical random mutagenesis and metabolic engineering to enhance FK506 production in Streptomyces sp. RM7011. Appl Microbiol Biotechnol 97:3053A3062

    Google Scholar 

  • Rajput KN, Patel KC, Trivedi UB (2016) Screening and selection of medium components for cyclodextrin glucanotransferase production by new Alkaliphile Microbacterium terrae KNR 9 using Plackett-Burman design. Biotechnol Res Int 2016:1–7

    Google Scholar 

  • Rakicka M, Rywińska A, Cybulski K, Rymowicz W (2016) Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants. Braz J Microbiol 2:417–423

    Article  Google Scholar 

  • Rocha VJ, Estrada M, Galindo E, Serrano CL (2006) From shake flasks to stirred fermentors: scale-up of an extractive fermentation process for 6-pentyl-α-pyrone production by trichoderma harzianum using volumetric power input. Process Biochem 41:1347–1352

    Article  Google Scholar 

  • Sheng L, Tang G, Su P, Zhang J, Xiao Q, Tong Q, Ma M (2016) Understanding the influence of tween 80 on pullulan fermentation by Aureobasidium pullulans CGMCC1234. Carbohyd Polym 136:1332–1337

    Article  CAS  Google Scholar 

  • Shirling EB, Gottlieb D (1966) Methods for characterization of Streptomyces species. Int J Syst Bacteriol 16:313–340

    Article  Google Scholar 

  • Singh V, Haque S, Niwas R, Srivastava A, Pasupuleti M, Tripathi CK (2017) Strategies for fermentation medium optimization: an in-depth review. Front Microbiol 7:2087

    Article  Google Scholar 

  • Srivastava A, Singh V, Tripath CKM (2018) Scale up and optimization of cholesterol oxidase production from Streptomyces rimosus MTCC 10792 in a 3-L bioreactor. Environ Sustain 1:99–107

    Article  Google Scholar 

  • Sun W, Zhou T, Xie H, Xiong Z (1994) Studies on the condition of fermentation of pullulan by Aureobasidium pullulans. Chin J Biotechnol 4:283–290

    Article  Google Scholar 

  • Talukdar S, Talukdar M, Buragohain M, Yadav A, Yadav RN, Bora TC (2016) Enhanced candicidal compound production by a new soil isolate Penicillium verruculosum MKH7 under submerged fermentation. BMC Microbiol 16:288

    Article  Google Scholar 

  • Taoka Y, Nagano N, Okita Y, Izumida H, Sugimoto S, Hayashi M (2011) Effect of tween 80 on the growth, lipid accumulation andfatty acid composition of Thraustochytrium aureum ATCC34304. J Biosci Bioeng 111:420–424

    Article  CAS  Google Scholar 

  • Turło J, Gajzlerska W, Klimaszewska M, Król M, Dawidowski M, Gutkowska B (2012) Enhancement of tacrolimus productivity in Streptomyces tsukubaensis by the use of novel precursors for biosynthesis. Enzyme Microb Technol 51:388–395

    Article  Google Scholar 

  • Verma T, Wei X, Lau SK, Bianchini A, Eskridge KM, Stratton J, Anderson NM, Thippareddi H, Subbiah J (2018) Response surface methodology for Salmonella inactivation during extrusion processing of oat flour. J Food Prot 81:815–826

    Article  Google Scholar 

  • Wang C, Liu J, Liu H, Liang S, Wen J (2017a) Combining metabolomics and network analysis to improve tacrolimus production in Streptomyces tsukubaensis using different exogenous feedings. J Ind Microbiol Biotechnol 11:11527–11540

    Google Scholar 

  • Wang J, Liu H, Huang D, Jin L, Wang C, Wen J (2017b) Comparative proteomic and metabolomic analysis of Streptomyces tsukubaensis reveals the metabolic mechanism of FK506 overproduction by feeding soybean oil. Appl Microbiol Biotechnol 6:2447–2465

    Article  Google Scholar 

  • Wang L, Zhao H, He D, Wu Y, Jin L, Li G, Su N, Li H, Xing XH (2020) Insights into the molecular-level effects of atmospheric and room-temperature plasma on mononucleotides and single-stranded homo- and hetero-oligonucleotides. Sci Rep 10(1):14298

    Article  CAS  Google Scholar 

  • Xia M, Huang D, Li S, Wen J, Jia X, Chen Y (2013) Enhanced FK506 production in Streptomyces tsukubaensis by rational feeding strategies based on comparative metabolic profiling analysis. Biotechnol Bioeng 110:2717–2730

    Article  CAS  Google Scholar 

  • Yang X, Xiao X, Liu D, Wu R, Wu C, Zhang J, Huang J, Liao B, He H (2017) Optimization of collagenase production by Pseudoalteromonas sp. sjn2 and application of collagenases in the preparation of antioxidative hydrolysates. Marine Drugs 15:377

    Article  Google Scholar 

  • Yoon YJ, Choi CY (1997) Nutrient effects on FK-506, a new immunosuppressant, production by Streptomyces sp. in a defined medium. J Ferment Bioeng 83:599–603

    Article  CAS  Google Scholar 

  • Zhang B, Cheung P (2011) A mechanistic study of the enhancing effect of tween 80 on the mycelial growth and exopolysaccharide production by Pleurotus tuber-regium. Bioresour Technol 17:8323

    Article  Google Scholar 

Download references

Funding

This work was supported by Special Project of Public Welfare Research Institute of Fujian Province of China (Grant Nos. 2019R1005-8 and 2020R1005009). The funders had no role in study design, data collection, and interpretation of results or manuscript writing.

Author information

Authors and Affiliations

  1. Fujian Provincial Key Laboratory of Screening for Novel Microbial Products, Fujian Institute of Microbiology, Fuzhou, 350007, China

    Lingbin Yan, Zhulan Zhang, Yin Zhang, Huangjian Yang, Guanrong Qiu, Desen Wang & Yunyang Lian

Authors
  1. Lingbin Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhulan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Huangjian Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Guanrong Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Desen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yunyang Lian
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

LY designed the research and prepared the manuscript. YL and ZZ supervised the research work and guided the experimental design. YZ, HY, DW and GQ provided the suggestion of the research work. All authors contributed to interpretation of the results and manuscript drafting. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Zhulan Zhang or Yunyang Lian.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yan, L., Zhang, Z., Zhang, Y. et al. Improvement of tacrolimus production in Streptomyces tsukubaensis by mutagenesis and optimization of fermentation medium using Plackett–Burman design combined with response surface methodology. Biotechnol Lett 43, 1765–1778 (2021). https://doi.org/10.1007/s10529-021-03144-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-021-03144-8

Keywords

Search

Navigation