Skip to main content
SpringerLink
Log in

Enhanced Production of Iturin A-2 Generated from Bacillus velezensis T701 and the Antitumor Activity of Iturin A-2 against Human Gastric Carcinoma Cells

  • Published:
International Journal of Peptide Research and Therapeutics Aims and scope Submit manuscript

Abstract

Bacillus velezensis T701 was used to produce lipopeptide. Among the lipopeptide, iturin A-2 was of the highest antitumor activity. In order to get higher yield of iturin A-2, the fermentation conditions of strain T701 were modified by optimization of medium composition and culture conditions. It was found that the concentrations of sucrose and peptone, and fermentation time had a very significant effect on the yield of iturin A-2 and fermentation temperature had a great influence on both of the yield of iturin A-2 and the proportion of iturin A-2 in iturin A. Another important conclusion was that monovalent cation could enhance metabolic ability of strain T701 and promote the yield of iturin A-2. Finally, under the optimized fermentation conditions of strain T701, iturin A-2 accounted for 80.2% of iturin A in HPLC trace, compared with the reported value of 46.8%. The yield of iturin A-2 increased to 1056.9 mg l−1. After separated and purified, pure iturin A-2 was achieved and confirmed by MS and NMR. The IC50 values of iturin A-2 against human gastric carcinoma cells NCI-N87 and MGC 803 were (51.7 ± 5.7) and (41.2 ± 4.3) μg ml−1, respectively. This is the first report on gram-scale preparation of iturin A-2 which also exhibits inhibitory activity against gastric cancer.

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

Similar content being viewed by others

References

  • Aftab U, Sajid I (2017) Antitumor peptides from Streptomyces sp. SSA 13, isolated from Arabian Sea. Int J Pept Res Ther 23:199–211

    Article  CAS  Google Scholar 

  • Ambrico A, Trupo M, Magarelli RA (2019) Influence of phenotypic dissociation in Bacillus subtilis strain ET-1 on iturin A production. Curr Microbiol 76(12):1487–1494

    Article  CAS  Google Scholar 

  • Chen W, Li X, Ma X, Chen S, Kang Y, Yang M, Huang F, Wan X (2019) Simultaneous hydrolysis with lipase and fermentation of rapeseed cake for iturin A production by Bacillus amyloliquefaciens CX-20. BMC Biotechnol 19:98

    Article  CAS  Google Scholar 

  • Dang Y, Zhao F, Liu X, Fan X, Huang R, Gao W, Wang S, Yang C (2019) Enhanced production of antifungal lipopeptide iturin A by Bacillus amyloliquefaciens LL3 through metabolic engineering and culture conditions optimization. Microb Cell Fact 18:68

    Article  Google Scholar 

  • Dey G, Bharti R, Dhanarajan G, Das S, Dey KK, Kumar BNP, Sen R, Mandal M (2015) Marine lipopeptide Iturin A inhibits Akt mediated GSK3β and FoxO3a signaling and triggers apoptosis in breast cancer. Sci Rep 5:10316

    Article  CAS  Google Scholar 

  • Dey G, Bharti R, Das AK, Sen R, Mandal M (2017a) Resensitization of Akt induced docetaxel resistance in breast cancer by ‘Iturin A’ a lipopeptide molecule from marine bacteria Bacillus megaterium. Sci Rep 7:17324

    Article  Google Scholar 

  • Dey G, Bharti R, Ojha PK, Pal I, Rajesh Y, Banerjee I, Banik P, Parida S, Parekh A, Sen R, Mandal M (2017b) Therapeutic implication of ‘Iturin A’ for targeting MD-2/TLR4 complex to overcome angiogenesis and invasion. Cell Signal 35:24–36

    Article  CAS  Google Scholar 

  • Ebe S, Ohike T, Matsukawa T, Okanami M, Kajiyama S, Ano T (2019) Promotion of lipopeptide antibiotic production by Bacillus sp. IA in the presence of rice husk biochar. J Pestic Sci 44:33–40

    Article  CAS  Google Scholar 

  • Hbid C, Jacques P, Razafindralambo H, Mpoyo MK, Meurice E, Paquot M, Thonart P (1996) Influence of the production of two lipopeptides, Iturin A and Surfactin S1, on oxygen transfer during Bacillus subtilis fermentation. Appl Biochem Biotechnol 57:571–579

    Article  Google Scholar 

  • Jacques P, Hbid C, Destain J, Razafindralambo H, Paquot M, Pauw ED, Thonart P (1999) Optimization of biosurfactant lipopeptide production from Bacillus subtilis S499 by Plackett-Burman design. Appl Biochem Biotechnol 77:223–233

    Article  Google Scholar 

  • Jiang J, Zhang H, Zhang C, Han M, Du J, Yang X, Li W (2021) Production, purification and characterization of ‘Iturin A-2’ a lipopeptide with antitumor activity from Chinese Sauerkraut Bacterium Bacillus velezensis T701. Int J Pept Res Ther. https://doi.org/10.1007/s10989-021-10241-9

    Article  Google Scholar 

  • Jin H, Li K, Niu Y, Guo M, Hu C, Chen S, Huang F (2015) Continuous enhancement of iturin A production by Bacillus subtilis with a stepwise two-stage glucose feeding strategy. BMC Biotechnol 15:53

    Article  Google Scholar 

  • Khan AW, Rahman MS, Ano T (2009) Application of malt residue in submerged fermentation of Bacillus subtilis. J Environ Sci 21:S33–S35

    Article  Google Scholar 

  • Liu Y, Ding S, Dietrich R, Mӓrtlbauer E, Zhu K (2017) A biosurfactant inspired heptapeptide with improved specificity to kill MRSA. Angew Chem Int Edit 129:1–5

    Article  Google Scholar 

  • Maryam JA, Ivica D, Petar R, Slaviša S, Gertrud EM (2019) Effect-directed screening of Bacillus lipopeptide extracts via hyphenated high-performance thin-layer chromatography. J Chromatogr A 1605:460366. https://doi.org/10.1016/j.chroma.2019.460366

    Article  CAS  Google Scholar 

  • Moore GE, Woods LK (1977) Culture media for human cells—RPMI 1603, RPMI 1634, RPMI 1640 and GEM 1717. Tissue Cult Assoc Man 3:503–509

    Article  Google Scholar 

  • Morton HJ (1970) A survey of commercially available tissue culture media. In Vitro 6(2):89–108

    Article  CAS  Google Scholar 

  • Phae CG, Shoda M (1991) Investigation of optimal conditions for foam separation of iturin, an antifungal peptide produced by Bacillus subtilis. J Ferment Bioeng 71:118–121. https://doi.org/10.1016/0922-338X(91)90235-9

    Article  CAS  Google Scholar 

  • Rahman MS, Ano T, Shoda M (2006) Second stage production of iturin A by induced germination of Bacillus subtilis RB14. J Biotechnol 125:513–515

    Article  CAS  Google Scholar 

  • Rahman MS, Ano T, Shoda M (2007) Biofilm fermentation of iturin A by a recombinant strain of Bacillus subtilis 168. J Biotechnol 127:503–507

    Article  CAS  Google Scholar 

  • Rangarajan V, Dhanarajan G, Kumar R, Sen R, Mandal M (2012) Time-dependent dosing of Fe2+ for improved lipopeptide production by marine Bacillus megaterium. J Chem Technol Biotechnol 87:1661–1669. https://doi.org/10.1002/jctb.3814

    Article  CAS  Google Scholar 

  • Shih IL, Lin CY, Wu JY, Hsieh C (2009) Production of antifungal lipopeptide from Bacillus subtilis in submerged fermentation using shake flask and fermentor. Korean J Chem Eng 26:1652–1661. https://doi.org/10.1007/s11814-009-0237-0

    Article  CAS  Google Scholar 

  • Sun D, Liao J, Sun L, Wang Y, Liu Y, Deng Q, Zhang N, Xu D, Fang Z, Wang W, Gooneratne R (2019) Effect of media and fermentation conditions on surfactin and iturin homologues produced by Bacillus natto NT-6: LC–MS analysis. AMB Express 9:120

    Article  Google Scholar 

  • Tsuchimori N, Ikegami S, Miyashiro S, Tsuji T, Kida T, Shibai H (1986) Isolation of iturin A-2 as an inhibitor of cytokinesis of starfish embryos. Comp Biochem Physiol Part C: Comp Pharmacol 84:381–384. https://doi.org/10.1016/0742-8413(86)90109-X

    Article  Google Scholar 

  • Wu JY, Liao JH, Shieh CJ, Hsieh FC, Liu YC (2018) Kinetic analysis on precursors for iturin A production from Bacillus amyloliquefaciens BPD1. J Biosci Bioeng 126:630–635

    Article  CAS  Google Scholar 

  • Xu Y, Cai D, Zhang H, Gao L, Yang Y, Gao J, Li Y, Yang C, Ji Z, Yu J, Chen S (2020) Enhanced production of iturin A in Bacillus amyloliquefaciens by genetic engineering and medium optimization. Process Biochem 90:50–57. https://doi.org/10.1016/j.procbio.2019.11.017

    Article  CAS  Google Scholar 

  • Ye Y, Li Q, Fu G, Yuan G, Miao J, Lin W (2012) Identification of antifungal substance (iturin A-2) produced by Bacillus subtilis B47 and its effect on southern corn leaf blight. J Integr Agric 11:90–99. https://doi.org/10.1016/S1671-2927(12)60786-X

    Article  CAS  Google Scholar 

  • Zhao H, Li J, Zhang Y, Lei S, Zhao X, Shao D, Jiang C, Shi J, Sun H (2018a) Potential of iturins as functional agents: safe, probiotic, and cytotoxic to cancer cells. Food Funct 9(11):5580–5587

    Article  CAS  Google Scholar 

  • Zhao H, Yan L, Xu X, Jiang C, Shi J, Zhang Y, Liu L, Lei S, Shao D, Huang Q (2018b) Potential of Bacillus subtilis lipopeptides in anti-cancer I: induction of apoptosis and paraptosis and inhibition of autophagy in K562 cells. AMB Express 8:78–93

    Article  Google Scholar 

  • Zhang Z, Ding ZT, Zhong J, Zhou JY, Shu D, Luo D, Yang J, Tan H (2017) Improvement of iturin A production in Bacillus subtilis ZK0 by overexpression of the comA and sigA genes. Lett Appl Microbiol 64:452–458

    Article  CAS  Google Scholar 

  • Zohora US, Rahman MS, Kahn AW, Okanami M, Ano T (2013) Improvement of production of lipopeptide antibiotic iturin A using fish protein. J Environ Sci 25:S2–S7

    Article  Google Scholar 

Download references

Acknowledgements

The authors acknowledge the financial support from the Natural Science Foundation of Hebei Province (CN) under Grant (Number B2016201031); and the Bureau of Science and Technology of Hebei Province (CN) under Grant (Numbers 20322901D and 21322802D).

Author information

Authors and Affiliations

  1. Key Laboratory of Chemical Biology of Hebei Province, Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding, 071002, People’s Republic of China

    Junpo Jiang, Mengnan Han, Shuangqing Fu, Jie Du, Honglei Zhang & Wei Li

  2. College of Life Science, Hebei Agricultural University, Baoding, 071001, People’s Republic of China

    Junpo Jiang & Shiying Wang

Authors
  1. Junpo Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mengnan Han
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shuangqing Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jie Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shiying Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Honglei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wei Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

JJ designed and performed experiments; WL and HZ supervised, and coordinated the study; MH, JD, SF and SW contributed to data analysis; JJ wrote the first draft manuscript; WL and HZ contributed to manuscript editing. All authors contributed substantially to this research and approved the final manuscript.

Corresponding authors

Correspondence to Honglei Zhang or Wei Li.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Ethical Approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher's Note

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

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOC 2176 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jiang, J., Han, M., Fu, S. et al. Enhanced Production of Iturin A-2 Generated from Bacillus velezensis T701 and the Antitumor Activity of Iturin A-2 against Human Gastric Carcinoma Cells. Int J Pept Res Ther 28, 27 (2022). https://doi.org/10.1007/s10989-021-10340-7

Download citation

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10989-021-10340-7

Keywords

Search

Navigation