盐酸胍/Ⅰ型胶原分散体系的流变性

doi:10.7503/cjcu20160063

摘要/Abstract

摘要：

采用胃蛋白酶降解法从猪皮中提取了胶原蛋白, 用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)确定为Ⅰ型胶原; 红外及紫外光谱表明胶原分子中存在三螺旋结构. 分别采用小幅振荡剪切法、恒定剪切速率法及滞后环法研究了盐酸胍浓度及作用时间对Ⅰ型胶原蛋白体系流变性的影响. 研究表明, 随盐酸胍浓度的增大及作用时间的延长, 盐酸胍/胶原分散体系由假塑性流体逐渐接近于牛顿流体. 在所研究的盐酸胍浓度范围(0~6.0 mol/L)内, 盐酸胍/胶原分散体系的触变性类型随盐酸胍浓度的增大发生正触变性-复合触变性-负触变性的转变; 随盐酸胍作用时间的延长(6~48 h), 盐酸胍浓度为1.0 mol/L的胶原分散体系的触变性类型发生复合触变性-负触变性的转变. 本文的研究结果有助于加深对触变性产生机理的认识.

关键词: 胶原, 盐酸胍, 变性, 流变性, 复合触变性

Abstract:

Collagen was extracted from the outer skin of pig using a pepsin digestion method. The isolated collagen was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE), Fourier transform infrared spectroscopy(FTIR) and ultraviolet(UV). According to the electrophoretic pattern, the isolated collagen consisted of two different α-chains(α₁, α₂) and one β-chain, indicating type Ⅰ collagen. FTIR and UV results confirmed the presence of triple helical structure in the collagen molecule. The rheological properties of GuHCl/type Ⅰ collagen dispersions were investigated as a function of guanidine hydrochloride(GuHCl) concentration and action time. The results of flow curves indicated that the pseudoplastic flow behavior of GuHCl/collagen dispersions gradually close to the Newtonian fluid with the increase of GuHCl concentration in the range of 0—6.0 mol/L and action time in the range of 6—48 h. The linear oscillatory shear, steady shear and thixotropic loop experiments were performed to investigate thixotropic behavior of the GuHCl/collagen dispersions. It was found that the thixotropic type of GuHCl/collagen dispersions transformed from positive thixotropy to complex thixotropy and then to negative thixotropy with the increase of GuHCl concentration. The thixotropic type of collagen dispersion transformed from complex thixotropy to negative thixotropy with the increase of action time of 1.0 mol/L GuHCl. It should be note that the complex thixotropy in GuHCl/collagen dispersions has not been reported in the similar systems so far. And we tentatively discussed this novel discovery on the basis of the interactions between GuHCl molecule and collagen. Meanwhile, this work would be helpful to deeply understand the mechanism of thixotropy.

Key words: Collagen, GuHCl, Denaturation, Rheological property, Complex thixotropy

中图分类号:

O631

TrendMD:

步红红, 焦燕妮, 兰文军, 庄晓丽, 戴肖南, 李天铎. 盐酸胍/Ⅰ型胶原分散体系的流变性. 高等学校化学学报, 2016, 37(6): 1189.

BU Honghong, JIAO Yanni, LAN Wenjun, ZHUANG Xiaoli, DAI Xiaonan, LI Tianduo. Rheological Properties of GuHCl/Type Ⅰ Collagen Dispersions^†. Chem. J. Chinese Universities, 2016, 37(6): 1189.

图/表 12

Fig.1 Protein patterns(A), FTIR(B) and UV-Vis(C) spectra of pig skin collagenLane 1 represents the protein molecular weight marker; Lane 2, 3 represent pig skin collagen.

Fig.2 SEM(A—D)and TEM(E, F) images of GuHCl/collagen dispersions(A) 0.1 mg/mL collagen; (B) 0.1 mg/mL collagen+0.5 mol/L GuHCl; (C) 0.1 mg/mL collagen+1.0 mol/L GuHCl;(D) 0.1 mg/mL collagen+4.0 mol/L GuHCl; (E) 0.1 mg/mL collagen; (F) 0.1 mg/mL collagen+4.0 mol/L GuHCl.

Fig.3 Flow curves of GuHCl/collagen dispersions at different GuHCl concentrations

Table 1 Power-law equation and R2 of GuHCl/collagen dispersions at different GuHCl concentrations

System	Power-law equation	R²
Collagen	τ=0.544 D^0.49	0.9957
Collagen+1.0 mol/L GuHCl	τ=0.073 D^0.68	0.9952
Collagen+2.0 mol/L GuHCl	τ=0.003 D^0.92	0.9961
Collagen+4.0 mol/L GuHCl	τ=0.002 D^0.95	0.9936
Collagen+6.0 mol/L GuHCl	τ=0.001 D^0.96	0.9967

Fig.4 Effect of 1.0 mol/L GuHCl function time on flow curves of collagen dispersionsTime/h: a. 6; b. 12; c. 24; d. 48.

Table 2 Power-law equation and R2 of GuHCl/collagen dispersions at different GuHCl(1.0 mol/L) function time

Time/h	Power-law equation	R²
6	τ=0.128 D^0.61	0.9946
12	τ=0.073 D^0.68	0.9952
24	τ=0.045 D^0.77	0.9969
48	τ=0.002 D^0.95	0.9980

Fig.5 Effect of GuHCl concentration on the thixotropy of collagen dispersions under oscillatory shear experimentsc(GuHCl)/(mol·L-1): (A) a. 0.25; b. 0.5; c. 0.75; d. 0. (B) a. 1.0; b. 2.0; c. 4.0; d. 6.0.

Fig.6 Effect of GuHCl function time on the thixotropy of collagen dispersions under oscillatory shear experiments

Fig.7 Thixotropy of collagen dispersions with different concertrations of GuHCl under steady shear experiments

Fig.8 Thixotropy of collagen dispersions with different function time of GuHCl under steady shear experiments

Fig.9 Thixotropic loop of collagen dispersions with different c(GuHCl)

Fig.10 Thixotropic loop of collagen dispersions with different function time of GuHCl(1.0 mol/L)Time/h: a. 6; b. 12; c. 24; d. 48.

参考文献 22

[1]	Doublet B., Van Der Rest M., J. Biol. Chem., 1991, 266, 6853—6858
[2]	Friess W., Eur. J. Pharm. Biopharm., 1998, 45, 113—136
[3]	Blair H. C., Zaidi M., Schlesinger P. H., Biochem. J., 2002, 354, 329—341
[4]	Brekken R. A., Sage E. H., Matrix Biology, 2000, 19, 569—580
[5]	Kadler K., Pretein Profile., 1995, 2, 491—519
[6]	Meaney M. M., Rice K., Wright R. J., Spector M., J. Orthop., Res., 2003, 21, 238—244
[7]	Usha R., Ramasami T., Themochimica Acta, 2004, 409, 201—206
[8]	Sionkowska A., Kaminska A., Inter. J. Biol. Macromolecules, 1999, 24, 337—340
[9]	McClain P. E., Wiley E. R., J. Biol. Chem., 1972, 247(3), 692—697
[10]	Wang F., Tang K. Y., Pan H. B., China Leather, 2011, 40(7), 30—33
	(王芳, 汤克勇, 潘洪波. 中国皮革, 2011, 40(7), 30—33)
[11]	Péterfi T., Arch. Entwicklungsmech. Organ., 1927, 112, 660—695(Péterfi T., Wilhelm Roux’ Archiv für Entwicklungsmechanik der Organismen, 1927,112, 660—695)
[12]	Dai X. N., Hou W. G., Duan H. D., Ni P., Colloid Surf. A, 2007, 295, 139—145
[13]	Chen Y., Ye R., Wang Y., Int. J. Food Sci. Tech., 2015, 50, 186—193
[14]	Zhang M., Liu W., Li G., Food Chemistry, 2009, 115, 826—831
[15]	Abraham L. C., Zuena E., Perez B., Kaplan D. L., Journal of Biomedical Materials Research B, 2008, 87, 264—285
[16]	Forgacs G., Newman S. A., Hinner B., Maier C. W., Sackmann E., Biophysical Journal, 2003, 84, 1272—1280
[17]	Kadler K. E., Hojima Y., Prockop D. J., J. Biol. Chem., 1987, 262, 15696—15701
[18]	Yan M. Y., Li B. F., Zhao X., Food Chemistry, 2010, 122, 1333—1337
[19]	Shi X. Y., Ma W. Y., Biomaterials, 2001, 22, 1627—1634
[20]	Wu K., Liu W. T., Li G. Y., Spectrochimica Acta A, 2013, 102, 186—193
[21]	Liu A. J., Chen Y., Jia Y. L., Zhang G. R., Process Biochemistry, 2007, 42, 54—59
[22]	Dai X. N., Hou W. G., Li S. P., Wang X. F., Chem. J. Chinese Universities, 2001, 22(9), 1578—1580
	(戴肖南, 侯万国, 李淑萍, 王新芳. 高等学校化学学报, 2001, 22(9), 1578—1580)