Processing and Modification of Recombinant Spider Silk Proteins_Journal of Biomedical Engineering

Authors：

LIUBin ¹ , WANGTao ² , LIUXiaobing ³ ,  LUOYougen ¹

1. School of Medicine, Jinggangshan University, Jian 343009, China;
2. School of Clinical Medicine, Jinggangshan University, Jian 343009, China;
3. School of Chemistry and Chemical Engineering, Jinggangshan University, Jian 343009, China;

Corresponding author：

LUOYougen, Email: lyougen@163.com

Keywords：

spider silk protein; recombinant spider silk protein; processing and modification; biomaterial

DOI：

10.7507/1001-5515.20150167

Video：

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Due to its special sequence structure, spider silk protein has unique physical and chemical properties, mechanical properties and excellent biological properties. With the expansion of the application value of spider silk in many fields as a functional material, progress has been made in the studies on the expression of recombinant spider silk proteins through many host systems by gene recombinant techniques. Recombinant spider silk proteins can be processed into high performance fibers, and a wide range of non-fibrous morphologies. Moreover, for their excellent biocompatibility and low immune response they are ideal for biomedical applications. Here we review the process and mechanism of preparation in vitro, chemistry and genetic engineering modification on recombinant spider silk protein.

Citation： LIUBin, WANGTao, LIUXiaobing, LUOYougen. Processing and Modification of Recombinant Spider Silk Proteins. Journal of Biomedical Engineering, 2015, 32(4): 933-939. doi: 10.7507/1001-5515.20150167 Copy

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2.	裔婷婷,潘志娟.静电纺再生加工对横纹金珠丝微观结构的影响[J].纺织学报,2012,33(4):1-5.
3.	CHUNG H,KIM T Y,LEE S Y.Recent advances in production of recombinant spider silk proteins[J].Curr Opin Biotechnol,2012,23(6):957-964.
4.	LEWICKA M,HERMANSON O,RISING A U.Recombinant spider silk matrices for neural stem cell cultures[J].Biomaterials,2012,33(31):7712-7717.
5.	赵亮,徐艳丽,邱慧,等.蛛丝蛋白双层小直径血管支架的制备及其血液相容性体外实验[J].中国修复重建外科杂志,2013,27(7):800-804.
6.	WIDHE M,JOHANSSON J,HEDHAMMAR M,et al.Current progress and limitations of spider silk for biomedical applications[J].Biopolymers,2012,97(6):468-478.
7.	HEIDEBRECHT A,SCHEIBEL T.Recombinant production of spider silk proteins[J].Adv Appl Microbiol,2013,82:115-153.
8.	GAO Zhenwei,LIN Zhi,HUANG Weidong,et al.Structural characterization of minor ampullate spidroin domains and their distinct roles in fibroin solubility and fiber formation[J].PLoS One,2013,8(2):e56142.
9.	王金雷,陈格飞,李佳,等.悦目金蛛丝蛋白TuSp1重复模块特征[J].东华大学学报:自然科学版,2013,39(2):196-201.
10.	PAQUET-MERCIER F,LEFEVRE T,AUGER M A.Evidence by infrared spectroscopy of the presence of two types of β-sheets in major ampullate spider silk and silkworm silk[J].Soft Matter,2013,9(1):208-215.
11.	杨子江,陈格飞,孟清.大腹园蛛次壶腹腺丝的表达[J].生物工程学报,2013,29(9):1323-1331.
12.	TEULÉ F,ADDISON B,COOPER A R,et al.Combining flagelliform and dragline spider silk motifs to produce tunable synthetic biopolymer fibers[J].Biopolymers,2012,97(6):418-431.
13.	RISING A,WIDHE M,JOHANSSON J,et al.Spider silk proteins:recent advances in recombinant production,structure-function relationships and biomedical applications[J].Cell Mol Life Sci,2011,68(2):169-184.
14.	WEN Hongxiu,LAN Xiqian,ZHANG Yuansong,et al.Transgenic silkworms (Bombyx mori) produce recombinant spider dragline silk in cocoons[J].Mol Biol Rep,2010,37(4):1815-1821.
15.	XIA Xiaoxia,QIAN Zhigang,KI C S,et al.Native-sized recombinant spider silk protein produced in metabolically engineered Escherichia coli results in a strong fiber[J].Proc Natl Acad Sci U S A,2010,107(32):14059-14063.
16.	TOKAREVA O,MICHALCZECHEN-LACERDA V A,RECH E L,et al.Recombinant DNA production of spider silk proteins[J].Microb Biotechnol,2013,6(6):651-663.
17.	HUMENIK M,SMITH A M,SCHEIBEL T.Recombinant spider silks-biopolymers with potential for future applications[J].Polymers (Basel),2011,3(1):640-661.
18.	ASKARIEH G,HEDHAMMAR M,NORDLING K,et al.Self-assembly of spider silk proteins is controlled by a pH-sensitive relay[J].Nature,2010,465(7295):236-238.
19.	HAGN F,EISOLDT L,HARDY J G,et al.A conserved spider silk domain acts as a molecular Switch that controls fibre assembly[J].Nature,2010,465(7295):239-242.
20.	SLOTTA U,MOUGIN N,ROEMER L,et al.Synthetic spider silk proteins and threads[J].Chemical Engineering Progress,2012,108(5):43-49.
21.	HU X,VASANTHAVADA K,KOHLER K,et al.Molecular mechanisms of spider silk[J].Cell Mol Life Sci,2006,63(17):1986-1999.
22.	RISING A,JOHANSSON J,LARSON G,et al.Major ampullate spidroins from Euprosthenops australis: multiplicity at protein,mRNA and gene levels[J].Insect Mol Biol,2007,5(16):551-561.
23.	KETEN S,XU Zhiping,IHLE B,et al.Nanoconfinement controls stiffness,strength and mechanical toughness of β-sheet crystals in silk[J].Nat Mater,2010,9(4):359-367.
24.	RAMMENSEE S,SLOTTA U,SCHEIBEL T,et al.Assembly mechanism of recombinant spider silk proteins[J].Proc Natl Acad Sci U S A,2008,105(18):6590-6595.
25.	HAGN F,THAMM C,SCHEIBEL T,et al.pH-dependent dimerization and salt-dependent stabilization of the N-terminal domain of spider dragline silk——implications for fiber formation[J].Angew Chem Int Ed Engl,2011,50(1):310-313.
26.	赵晓,张袁松,曾峥,等.基于天然蚕丝及蜘蛛丝蛋白的生物材料研究进展[J].蚕学通讯,2010,30(2):31-37.
27.	RABOTYAGOVA O S,CEBE P,KAPLAN D L.Self-assembly of genetically engineered spider silk block copolymers[J].Biomacromolecules,2009,10(2):229-236.
28.	LAMMEL A,SCHWAB M,SLOTTA U,et al.Processing conditions for the formation of spider silk microspheres[J].ChemSusChem,2008,1(5):413-416.
29.	LAMMEL A,SCHWAB M,HOFER M,et al.Recombinant spider silk particles as drug delivery vehicles[J].Biomaterials,2011,32(8):2233-2240.
30.	HOFER M,WINTER G,MYSCHIK J.Recombinant spider silk particles for controlled delivery of protein drugs[J].Biomaterials,2012,33(5):1554-1562.
31.	LIEBMANN B,HVMMERICH D,SCHEIBEL T,et al.Formulation of poorly water-soluble substances using self-assembling spider silk protein[J].Colloids Surf A Physicochem Eng Asp,2008,331(1/2):126-132.
32.	YOUNG S L,GUPTA M,HANSKE C,et al.Utilizing conformational changes for patterning thin films of recombinant spider silk proteins[J].Biomacromolecules,2012,13(10):3189-3199.
33.	SPIESS K,LAMMEL A,SCHEIBEL T.Recombinant spider silk proteins for applications in biomaterials[J].Macromol Biosci,2010,10(9):998-1007.
34.	RENAULT A,RIOUX-DUBÉ J F,LEFÈVRE T,et al.Structure and mechanical properties of spider silk films at the air-water interface[J].Langmuir,2013,29(25):7931-7938.
35.	HORINEK D,SERR A,GEISLER M,et al.Peptide adsorption on a hydrophobic surface results from an interplay of solvation,surface,and intrapeptide forces[J].Proc Natl Acad Sci U S A,2008,105(8):2842-2847.
36.	HERMANSON K D,HUEMMERICH D,SCHEIBEL T A.Engineered microcapsules fabricated from reconstituted spider silk[J].Adv Mater,2007,19(14):1810.
37.	EISOLDT L,SMITH A,SCHEIBEL T.Decoding the secrets of spider silk[J].Materialstoday,2011,14(3):80-86.
38.	SLOTTA U,HESS S,SPIESS K,et al.Spider silk and amyloid fibrils:a structural comparison[J].Macromol Biosci,2007,7(2):183-188.
39.	RAMMENSEE S,HUEMMERICH D,HERMANSON K D,et al.Rheological characterization of hydrogels formed by recombinantly produced spider silk[J].Appl Phys A Mater Sci Process,2006,82(2):261-264.
40.	LEAL-EGAÑA A,SCHEIBEL T.Silk-based materials for biomedical applications[J].Biotechnol Appl Biochem,2010,55(3):155-167.
41.	SCHACHT K,SCHEIBEL T.Controlled hydrogel formation of a recombinant spider silk protein[J].Biomacromolecules,2011,12(7):2488-2495.
42.	HUEMMERICH D,SLOTTA U,SCHEIBEL T.Processing and modification of films made from recombinant spider silk proteins[J].Appl Phys A Mater Sci Process,2006,82(2):219-222.
43.	SPIESS K,WOHLRAB S,SCHEIBEL T.Structural characterization and functionalization of engineered spider silk films[J].Soft Matter,2010,6(17):4168-4174.
44.	WOHLRAB S,MVLLER S,SCHMIDT A,et al.Cell adhesion and proliferation on RGD-modified recombinant spider silk proteins[J].Biomaterials,2012,33(28):6650-6659.
45.	TEULÉ F,MIAO Yungen,SOHN B H,et al.Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties[J].Proc Natl Acad Sci U S A,2012,109(3):923-928.
46.	HAUPTMANN V,WEICHERT N,MENZEL M,et al.Native-sized spider silk proteins synthesized in planta via intein-based multimerization[J].Transgenic Res,2013,22(2):369-377.
47.	HUANG Huahua,HU Jinlian,ZHU Yong.Shape-memory biopolymers based on β-sheet structures of polyalanine segments inspired by spider silks[J].Macromol Biosci,2013,13(2):161-166.
48.	CURRIE H A,DESCHAUME O,NAIK R R,et al.Genetically engineered chimeric silk-silver binding proteins[J].Adv Funct Mater,2011,21(15):2889-2895.
49.	KRISHNAJI S T,KAPLAN D L.Bioengineered chimeric spider silk-uranium binding proteins[J].Macromol Biosci,2013,13(2):256-264.
50.	刘全勇,江雷.仿生学与天然蜘蛛丝仿生材料[J].高等学校化学学报,2010,31(6):1065-1071.

1. CRANFORD S W,TARAKANOVA A,PUGNO N M,et al.Nonlinear material behaviour of spider silk yields robust webs[J].Nature,2012,482(7383):72-76.
2. 裔婷婷,潘志娟.静电纺再生加工对横纹金珠丝微观结构的影响[J].纺织学报,2012,33(4):1-5.
3. CHUNG H,KIM T Y,LEE S Y.Recent advances in production of recombinant spider silk proteins[J].Curr Opin Biotechnol,2012,23(6):957-964.
4. LEWICKA M,HERMANSON O,RISING A U.Recombinant spider silk matrices for neural stem cell cultures[J].Biomaterials,2012,33(31):7712-7717.
5. 赵亮,徐艳丽,邱慧,等.蛛丝蛋白双层小直径血管支架的制备及其血液相容性体外实验[J].中国修复重建外科杂志,2013,27(7):800-804.
6. WIDHE M,JOHANSSON J,HEDHAMMAR M,et al.Current progress and limitations of spider silk for biomedical applications[J].Biopolymers,2012,97(6):468-478.
7. HEIDEBRECHT A,SCHEIBEL T.Recombinant production of spider silk proteins[J].Adv Appl Microbiol,2013,82:115-153.
8. GAO Zhenwei,LIN Zhi,HUANG Weidong,et al.Structural characterization of minor ampullate spidroin domains and their distinct roles in fibroin solubility and fiber formation[J].PLoS One,2013,8(2):e56142.
9. 王金雷,陈格飞,李佳,等.悦目金蛛丝蛋白TuSp1重复模块特征[J].东华大学学报:自然科学版,2013,39(2):196-201.
10. PAQUET-MERCIER F,LEFEVRE T,AUGER M A.Evidence by infrared spectroscopy of the presence of two types of β-sheets in major ampullate spider silk and silkworm silk[J].Soft Matter,2013,9(1):208-215.
11. 杨子江,陈格飞,孟清.大腹园蛛次壶腹腺丝的表达[J].生物工程学报,2013,29(9):1323-1331.
12. TEULÉ F,ADDISON B,COOPER A R,et al.Combining flagelliform and dragline spider silk motifs to produce tunable synthetic biopolymer fibers[J].Biopolymers,2012,97(6):418-431.
13. RISING A,WIDHE M,JOHANSSON J,et al.Spider silk proteins:recent advances in recombinant production,structure-function relationships and biomedical applications[J].Cell Mol Life Sci,2011,68(2):169-184.
14. WEN Hongxiu,LAN Xiqian,ZHANG Yuansong,et al.Transgenic silkworms (Bombyx mori) produce recombinant spider dragline silk in cocoons[J].Mol Biol Rep,2010,37(4):1815-1821.
15. XIA Xiaoxia,QIAN Zhigang,KI C S,et al.Native-sized recombinant spider silk protein produced in metabolically engineered Escherichia coli results in a strong fiber[J].Proc Natl Acad Sci U S A,2010,107(32):14059-14063.
16. TOKAREVA O,MICHALCZECHEN-LACERDA V A,RECH E L,et al.Recombinant DNA production of spider silk proteins[J].Microb Biotechnol,2013,6(6):651-663.
17. HUMENIK M,SMITH A M,SCHEIBEL T.Recombinant spider silks-biopolymers with potential for future applications[J].Polymers (Basel),2011,3(1):640-661.
18. ASKARIEH G,HEDHAMMAR M,NORDLING K,et al.Self-assembly of spider silk proteins is controlled by a pH-sensitive relay[J].Nature,2010,465(7295):236-238.
19. HAGN F,EISOLDT L,HARDY J G,et al.A conserved spider silk domain acts as a molecular Switch that controls fibre assembly[J].Nature,2010,465(7295):239-242.
20. SLOTTA U,MOUGIN N,ROEMER L,et al.Synthetic spider silk proteins and threads[J].Chemical Engineering Progress,2012,108(5):43-49.
21. HU X,VASANTHAVADA K,KOHLER K,et al.Molecular mechanisms of spider silk[J].Cell Mol Life Sci,2006,63(17):1986-1999.
22. RISING A,JOHANSSON J,LARSON G,et al.Major ampullate spidroins from Euprosthenops australis: multiplicity at protein,mRNA and gene levels[J].Insect Mol Biol,2007,5(16):551-561.
23. KETEN S,XU Zhiping,IHLE B,et al.Nanoconfinement controls stiffness,strength and mechanical toughness of β-sheet crystals in silk[J].Nat Mater,2010,9(4):359-367.
24. RAMMENSEE S,SLOTTA U,SCHEIBEL T,et al.Assembly mechanism of recombinant spider silk proteins[J].Proc Natl Acad Sci U S A,2008,105(18):6590-6595.
25. HAGN F,THAMM C,SCHEIBEL T,et al.pH-dependent dimerization and salt-dependent stabilization of the N-terminal domain of spider dragline silk——implications for fiber formation[J].Angew Chem Int Ed Engl,2011,50(1):310-313.
26. 赵晓,张袁松,曾峥,等.基于天然蚕丝及蜘蛛丝蛋白的生物材料研究进展[J].蚕学通讯,2010,30(2):31-37.
27. RABOTYAGOVA O S,CEBE P,KAPLAN D L.Self-assembly of genetically engineered spider silk block copolymers[J].Biomacromolecules,2009,10(2):229-236.
28. LAMMEL A,SCHWAB M,SLOTTA U,et al.Processing conditions for the formation of spider silk microspheres[J].ChemSusChem,2008,1(5):413-416.
29. LAMMEL A,SCHWAB M,HOFER M,et al.Recombinant spider silk particles as drug delivery vehicles[J].Biomaterials,2011,32(8):2233-2240.
30. HOFER M,WINTER G,MYSCHIK J.Recombinant spider silk particles for controlled delivery of protein drugs[J].Biomaterials,2012,33(5):1554-1562.
31. LIEBMANN B,HVMMERICH D,SCHEIBEL T,et al.Formulation of poorly water-soluble substances using self-assembling spider silk protein[J].Colloids Surf A Physicochem Eng Asp,2008,331(1/2):126-132.
32. YOUNG S L,GUPTA M,HANSKE C,et al.Utilizing conformational changes for patterning thin films of recombinant spider silk proteins[J].Biomacromolecules,2012,13(10):3189-3199.
33. SPIESS K,LAMMEL A,SCHEIBEL T.Recombinant spider silk proteins for applications in biomaterials[J].Macromol Biosci,2010,10(9):998-1007.
34. RENAULT A,RIOUX-DUBÉ J F,LEFÈVRE T,et al.Structure and mechanical properties of spider silk films at the air-water interface[J].Langmuir,2013,29(25):7931-7938.
35. HORINEK D,SERR A,GEISLER M,et al.Peptide adsorption on a hydrophobic surface results from an interplay of solvation,surface,and intrapeptide forces[J].Proc Natl Acad Sci U S A,2008,105(8):2842-2847.
36. HERMANSON K D,HUEMMERICH D,SCHEIBEL T A.Engineered microcapsules fabricated from reconstituted spider silk[J].Adv Mater,2007,19(14):1810.
37. EISOLDT L,SMITH A,SCHEIBEL T.Decoding the secrets of spider silk[J].Materialstoday,2011,14(3):80-86.
38. SLOTTA U,HESS S,SPIESS K,et al.Spider silk and amyloid fibrils:a structural comparison[J].Macromol Biosci,2007,7(2):183-188.
39. RAMMENSEE S,HUEMMERICH D,HERMANSON K D,et al.Rheological characterization of hydrogels formed by recombinantly produced spider silk[J].Appl Phys A Mater Sci Process,2006,82(2):261-264.
40. LEAL-EGAÑA A,SCHEIBEL T.Silk-based materials for biomedical applications[J].Biotechnol Appl Biochem,2010,55(3):155-167.
41. SCHACHT K,SCHEIBEL T.Controlled hydrogel formation of a recombinant spider silk protein[J].Biomacromolecules,2011,12(7):2488-2495.
42. HUEMMERICH D,SLOTTA U,SCHEIBEL T.Processing and modification of films made from recombinant spider silk proteins[J].Appl Phys A Mater Sci Process,2006,82(2):219-222.
43. SPIESS K,WOHLRAB S,SCHEIBEL T.Structural characterization and functionalization of engineered spider silk films[J].Soft Matter,2010,6(17):4168-4174.
44. WOHLRAB S,MVLLER S,SCHMIDT A,et al.Cell adhesion and proliferation on RGD-modified recombinant spider silk proteins[J].Biomaterials,2012,33(28):6650-6659.
45. TEULÉ F,MIAO Yungen,SOHN B H,et al.Silkworms transformed with chimeric silkworm/spider silk genes spin composite silk fibers with improved mechanical properties[J].Proc Natl Acad Sci U S A,2012,109(3):923-928.
46. HAUPTMANN V,WEICHERT N,MENZEL M,et al.Native-sized spider silk proteins synthesized in planta via intein-based multimerization[J].Transgenic Res,2013,22(2):369-377.
47. HUANG Huahua,HU Jinlian,ZHU Yong.Shape-memory biopolymers based on β-sheet structures of polyalanine segments inspired by spider silks[J].Macromol Biosci,2013,13(2):161-166.
48. CURRIE H A,DESCHAUME O,NAIK R R,et al.Genetically engineered chimeric silk-silver binding proteins[J].Adv Funct Mater,2011,21(15):2889-2895.
49. KRISHNAJI S T,KAPLAN D L.Bioengineered chimeric spider silk-uranium binding proteins[J].Macromol Biosci,2013,13(2):256-264.
50. 刘全勇,江雷.仿生学与天然蜘蛛丝仿生材料[J].高等学校化学学报,2010,31(6):1065-1071.

Journal of Biomedical Engineering

Processing and Modification of Recombinant Spider Silk Proteins

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