Allophycocyanin is one of the most important marine active peptides. Previous studies suggested that recombinant allophycocyanin (rAPC) could remarkably inhibit the S-180 carcinoma in mice, indicating its potential pharmaceutical uses. Based on intergeneric conjugal transfer, heterologous expression of rAPC was first achieved in marine Streptomyces sp. isolate M097 through inserting the apc gene into the thiostrepton-induced vector pIJ8600. The transformation frequency for this system was approximately 10−4 exconjugants/recipient. In the transformed Streptomyces sp. isolate M097, the yield of purified rAPC could amount to about 38 mg/l using a simple purification protocol, and HPLC analysis showed that the purity of the protein reached about 91.5%. In vitro activity tests also revealed that the purified rAPC had effective scavenging abilities on superoxide and hydroxyl radicals. This would widen the usefulness of the marine Streptomyces as a host to express the rAPC and to offer industrial strain for the production of rAPC.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Chiang S.J., 2004 Strain improvement for fermentation and biocatalysis processes by genetic engineering technology Journal of Industrial Microbiology and Biotechnology 31:99–108
Faury D., Saidane S., Li H., Morosoli R., 2004 Secretion of active xylanase C from Streptomyces lividans is exclusively mediated by the Tat protein export system Biochimica et Biophysica Acta. 1699:155–162
Flett F., Mersinias V., Smith C.P., 1997 High efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting Streptomyces FEMS Microbiology Letters 155:223–229
Fukatsu H., Herai S., Hashimoto Y., Maseda H., Higashibata H., Kobayashi M., 2005 High-level expression of a novel amine-synthesizing enzyme, N-substituted formamide deformylase, in Streptomyces with a strong protein expression system Protein Expression and Purification 40:212–219
Ge, B.S., Tang, Z.H., Zhao, F.Q., Ren, Y.H., Yang, Y. & Qin, S. 2005 Scale-up of the fermentation and purification of the recombinant allophycocyanin over-expressed in Escherichia coli. Process Biochemistry 40, 3190–3195
Glazer A.N., 1994 Phycobiliproteins-a family of valuably, widely used fluorophores Journal of Applied Phycology 6:105–112
Isiegas C., Parro V., Mellado R.P., 1999 Streptomyces lividans as a host for the production and secretion of Escherichia coli TEM ß-lactamase Letters in Applied Microbiology 28:321–326
Kieser T., Bibb M.J., Buttner M.J., Chater K.F., Hopwood D.A., 2000 Practical Streptomyces Genetics John Innes Foundation, Norwich ISBN 0-7084-0623-8
Kronick M.N., 1986 The use of phycobiliproteins as fluorescent labels in immunoassay Journal of Immunological Methods 92:1–13
Lambalot R.H., Cane D.E., 1992 Isolation and characterization of 10-deoxymethynolide produced by Streptomyces venezuelae Journal of Antibiotics 45:1981–1982
Li F.C., Maskey R.P., Qin S., Sattler I., Fiebig H.H., Maier A., Zeeck A., Laatsch H., 2005 Chinikomycins A and B: Isolation, structure elucidation, and biological activity of novel antibiotics from a marine Streptomyces sp. isolate M045 Journal of Natural Products 68:349–353
MacColl R., 2004 Allophycocyanin and energy transfer Biochimica et Biophysica Acta 1657:73–81
Nancy D.C., 2001 Expression systems for use in actinomycetes and related organisms Current Opinion in Biotechnology 12:446–449
Qin S., Tang Z.H., Lin F., Sung L.A., Tseng C.K., 2004 Genomic cloning, expression and recombinant protein purification of α and β subunits of the allophycocyanin gene (apc) from the cyanobacterium Anacystis nidulans UTEX 625 Journal of Applied Phycology 16:483–487
Schaerlaekens K., Lammertynm E., Geukensm N., Keersmaeker S.D., Anné J., Mellaert L.V., 2004 Comparison of the Sec and Tat secretion pathways for heterologous protein production by Streptomyces lividans Journal of Biotechnology 112:279–288
Storf M., Parbel A., Meyer M., Strohmann B., Scheer H., Deng M., Zheng M., Zhou M., Zhao K., 2001 Chromophore attachment to biliproteins: Specificity of PecE/PecF, a lyase/isomerase for the photoactive31-Cys-a84-phycoviolobilin chromophore of phycoerythrocyanin Biochemistry 40:12444–12456
Sun L., Wang S.M., 2003 Allophycocyanin complexes from the phycobilisome of a thermophilic blue-green alga Myxosarcina concinna Printz Journal of Photochemistry and Photobiology B: Biology 72:45–53
Tang S.M., Huang F., Dou C.G., Zhuang G.V., Qin S., Wang X.H., Zou L.H., 1996 The inhibition effect of recombinant allophycocyanin on S-180 carcinoma in mice Pharmaceutical Biotechnology 3:168–170
Tang Z.H., Zhao F.Q., Qin S., Wu S.J., Gao J.T., Ding J., 2004 The Production of rAPC and the study of its inhibition effect on H22 tumor High Technology Letters 3:83–86
Trinquet E., Maurin F., Preaudat M., Mathis G., 2001 Allophycocyanin I as a near-infrared fluorescent tracer: isolation, characterization, chemical modification, and use in a homogeneous fluorescence resonance energy transfer system Analytical Biochemistry 296:232–244
Zhang E.X., Yu L.J., Xiao X., 1995 Studies on oxygen free radical-scavenging effect of polysaccharide from Sargassum thunbergii Chinese Journal of Marine Drugs 53:1–4
Zhang Y., Wang W.C., Li Y., 2004 Cloning, expression, and purification of soluble human interleukin-4 receptor in Streptomyces Protein Expression and Purification 36:139–145
Zhao F.Q., Tang Z.H., Lin F., Qin S., Dou C.G., 2003 High cell density fermentation of a recombinant E. coli strain P1 producing an anti-S180 tumor recombinant protein (rAPC) High Technology Letters 2:29–32
We thank Prof Keith F. Chater, John Innes Institute, for the gift of the strain ET12567 (PUZ8002) and plasmid pIJ8600. This work was supported by National High Technology Research and Development Program of China (863 Program, 2003AA624020) and Key Innovative Project (KSCX2-SW-215) of the Chinese Academy of Sciences.
About this article
Cite this article
Hou, Y., Qin, S., Li, Y. et al. Heterologous Expression and Purification of Recombinant Allophycocyanin in Marine Streptomyces sp. Isolate M097. World J Microbiol Biotechnol 22, 525–529 (2006). https://doi.org/10.1007/s11274-005-9067-3
- conjugal transfer
- heterologous expression