Effects of Cryo-grinding on the Extraction and Physicochemical Properties of Silk Fibroin
-
摘要: 本文将冷冻研磨技术引入到丝素蛋白的提取工艺中，以期实现丝素蛋白高效制备且性能可控的目的。具体地，将蚕茧在液氮中预冻12 h后进行低温研磨，并将得到的样品（GSF322）与未研磨样品SF（对照）进行理化性能对比。结果表明，SF，GSF322的得率相当，但脱胶时间从90 min减少到40 min；圆二色谱分析显示，预处理过程不会破坏丝素蛋白的立体结构；浊度实验研究表明，通过调控研磨时间可以有效调节丝素蛋白的体外组装能力，研磨3 min的丝素样品具有最好的体外组装能力；Zeta电位实验结果显示，预处理后，GSF322分子链上带正电荷的基团增多，更多-NH2的暴露导致材料亲水性增强。溶解性实验则证明，经过研磨处理后的GSF322在水中溶解率更高。热稳定性测试中，GSF322的脱水峰较SF的峰值高，表明其热稳定性更好。综合来说，在丝素蛋白的提取中，用冷冻研磨法来进行蚕茧的预处理，不仅可以提高丝素的提取效率，而且也可以调控其理化性能。Abstract: Herein, cryo-grinding technique was introduced into the extraction process of silk fibroin, aiming to achieve efficient preparation and controllable performance of silk fibroin. More specifically, the cocoon was pre-frozen in liquid nitrogen for 12 h before being ground, and the physical and chemical properties of the obtained sample (GSF322) were compared with the control one (SF). Results revealed that, compared with SF, the degumming duration for GSF322 dwindled from 90 min to 40 min with a close yield. Circular dichroism analysis demonstrated that the pretreatment process would not destroy the steric structure of the macromolecule. Turbidity assay implied that the assembly activity of the polymer in vitro could also be manipulated via assorted grinding periods, among which the sample ground for 3 min had the best assembly capacity. Dissection of the Zeta potential displayed that, after grinding, the positively-charged groups on the molecular chain of GSF322 might multiply, resulting in the exposure of more -NH2 groups in the surroundings and enhanced hydrophilicity of the material, which was in accordance with the trend from the water solubility experiment. Moreover, in the thermostability test, the dehydration peak for GSF322 was higher than that of SF, showing better thermal stability. To conclude, concerning the extraction process for silk fibroin, the cryogrinding approach sheds light on both the strengthening of the extraction efficiency as well as the tuning of the physiochemical aptitudes of the polymer.
-
Key words:
- silk fibroin /
- degumming /
- cryo-grinding /
- self-assembly /
- thermal stability
-
图 1 不同冷冻研磨条件的再生丝素蛋白得率
Figure 1. Yield of regenerated silk fibroin protein under different freeze grinding conditions
图 3 不同冷冻研磨条件下丝素蛋白圆二色谱图
Figure 3. Circle chromatogram of silk fibroin under different freeze grinding conditions
图 4 不同冷冻研磨条件再生丝素蛋白浊度曲线图
Figure 4. Turbidity curves of regenerated silk fibroin protein under different freeze grinding conditions
-
[1] HUANG X W, LIANG H, LI Z, et a1. Monodisperse phase transfer and surface bioengineering of metal nanoparticles via a silk fibroin protein corona[J]. Nanoscale,2017,9(8):2695−2700. doi: 10.1039/C6NR09581J [2] LIM H R, KIM H S, QAZI R, et al. Advanced soft materials, sensor integrations, and applications of wearable flexible hybrid electronics in healthcare, energy, and environment[J]. Advanced Materials,2020,32:1901924.1−1901924.43. [3] ZHANG X H, SHENG N N, WANG L A, et al. Supramolecular nanofibrillar hydrogels as highly stretchable, elastic and sensitive ionic sensors[J]. Materials Horizons Journal,2019,6:326−333. doi: 10.1039/C8MH01188E [4] CHEN F, LU S P, ZHU L, et al. Conductive regenerated silk-fibroin-based hydrogels with integrated high mechanical performances[J]. J Mater Chem B,2019,7:1708−1715. doi: 10.1039/C8TB02445F [5] LI Y L, XIAO Y, LIU C S. The horizon of materiobiology: A perspective on material-guided cell behaviors and tissue engineering[J]. Chem Rev,2017,117:4376−4421. [6] DHASMANA A, SINGH L, ROY P, et al. Silk fibroin protein modified acellular dermal matrix for tissue repairing and regeneration[J]. Mat Sci Eng C-Mater Biol Appl,2019,97:313−324. doi: 10.1016/j.msec.2018.12.038 [7] 李平, 宛晓春, 陶文沂, 等. 丝素蛋白膜固定β葡萄糖苷酶及其改良食品风味的研究[J]. 菌物学报,2014,23(1):73−78. [LI P, WAN X C, TAO W Y, et al. Study on the immobilization of β-glucosidase by silk fibroin membrane and its improvement of food flavor[J]. Acta Bacteriologica Sinica,2014,23(1):73−78.LI P, WAN X C, TAO W Y, et al. Study on the immobilization of β-glucosidase by silk fibroin membrane and its improvement of food flavor[J]. Acta Bacteriologica Sinica, 2014, 23(1): 73-78. [8] MARELLI B, BRENCKLE M A, KAPLAN D L, et al. Silk fibroin as edible coating for perishable food preservation[J]. Scientific Reports,2016,6:25236. doi: 10.1038/srep25263 [9] 屠洁, 刘冠卉, 燕薇. 4种常用稳定剂和丝素蛋白对搅拌型酸奶黏度和保水性的影响[J]. 食品科学,2012,33(21):136−140. [TU J, LIU G H, YAN W. Effects of four common stabilizers combined with silk fibroin on viscosity and water-holding capacity of stirred yogurt[J]. Food Science,2012,33(21):136−140.TU J, LIU G H, YAN W. Effects of four common stabilizers combined with silk fibroin on viscosity and water-holding capacity of stirred yogurt[J]. Food Science, 2012, 33(21): 136-140. [10] 李莹莹, 王昉, 刘其春. 丝素蛋白及其复合材料的研究进展[J]. 材料工程,2018,46(8):14−26. [LI Y Y, WANG F, LIU Q C. Research progress of silk fibroin and its composite materials[J]. Materials Engineering,2018,46(8):14−26. doi: 10.11868/j.issn.1001-4381.2017.001242LI Y Y, WANG F, LIU Q C. Research progress of silk fibroin and its composite materials[J]. Materials Engineering, 2018, 46(8): 14-26. doi: 10.11868/j.issn.1001-4381.2017.001242 [11] ARANGO M C, MONTOYA Y, PERESIN M S, et al. Silk sericin as a biomaterial for tissue engineering: A review[J]. International Journal of Polymeric Materials,2021,70(15/18):1115−1129. [12] CHELAZZI D, BADILLO-SANCHEZ D, GIORGI R, et al. Self-regenerated silk fibroin with controlled crystallinity for the reinforcement of silk[J]. Journal of Colloid and Interface Science,2020,576:230−240. doi: 10.1016/j.jcis.2020.04.114 [13] GUO X, LIN N, LU S, et al. Preparation and biocompatibility characterization of silk fibroin 3D scaffolds[J]. ACS Applied Bio Materials,2021,4(2):1369−1380. doi: 10.1021/acsabm.0c01239 [14] JMA B, SM C, SG C, et al. Silk fibroin as biomaterial for bone tissue engineering[J]. Acta Biomaterialia,2016,31:1−16. doi: 10.1016/j.actbio.2015.09.005 [15] GUAN Y, YANG X Y, WANG L, et al. A novel silk/polyester woven small diameter arterial prosthesis: Degumming and the influence on cytocompatibility[J]. Fibers and Polymers,2015,16:1533−1539. doi: 10.1007/s12221-015-4934-5 [16] SUN W, GREGIRY D A, TOMEH M A, et al. Silk fibroin as a functional biomaterial for tissue engineering[J]. International Journal of Molecular Sciences,2021,22(3):1499. doi: 10.3390/ijms22031499 [17] WANG L P, LUO Z W, et al. Effect of degumming methods on the degradation behavior of silk fibroin biomaterials[J]. Fibers and Polymers,2019,20:45−50. doi: 10.1007/s12221-019-8658-9 [18] ZHENG Z, GUO S, LIU Y, et al. Lithium-free processing of silk fibroin[J]. Journal of Biomaterials Applications,2016,31(3):450−463. doi: 10.1177/0885328216653259 [19] RUI W, ZHU Y, ZHUO S, et al. Degumming of raw silk via steam treatment[J]. Journal of Cleaner Production,2018,203(PT.1−1216):492−497. [20] WANG Z, YANG H, LI W, et al. Effect of silk degumming on the structure and properties of silk fibroin[J]. The Journal of the Textile Institute,2019,110(1):134−140. doi: 10.1080/00405000.2018.1473074 [21] VYAS S K, SHUKLA S R. Comparative study of degumming of silk varieties by different techniques[J]. Journal of the Textile Institute Proceedings & Abstracts,2015,107(2):191−199. [22] RASTOGI S, KANDASU B. Processing trends of silk fibers: Silk degumming, regeneration and physical functionalization[J]. Journal of the Textile Institute,2020,111(12):1−17. [23] TOWNSEND B, PEYRONEL F, CAHHAGHAN-PATEACHAR N, et al. Shear-induced aggregation or disaggregation in edible oils: Models, computer simulation, and USAXS measurements[J]. J Appl Phys,2017,122(22):224304−224314. doi: 10.1063/1.5004023 [24] WILSON J F, BOLESLAV Z, ONDŘEJ Š, et al. Study of the shear-thinning effect between polymer nanoparticle surfaces during shear-induced aggregation[J]. Ind Eng Chem Res,2021,60(29):10654−10665. doi: 10.1021/acs.iecr.1c00232 [25] MEGHWAL M, GOSWAMI T K. Comparative study on ambient and cryogenic grinding of fenugreek and black pepper seeds using rotor, ball, hammer and pin mill[J]. Powder Technology,2014,267:245−255. doi: 10.1016/j.powtec.2014.07.025 [26] HE L, YANG J, XU C Z, et al. Effect of pre-shearing treatment on the molecular structure, fibrillogenesis behavior and gel properties of collagen[J]. New J Chem,2020,44(17):6760−6770. doi: 10.1039/D0NJ00054J [27] SUN M, WEI X, WANG H B, et al. Structure restoration of thermally denatured collagen by ultrahigh pressure treatment[J]. Food Bioprocess Tech,2020,13(2):367−378. doi: 10.1007/s11947-019-02389-6 [28] YANG J, WANG H B, HE L, et al. Reconstituted fibril from heterogenic collagens-a new method to regulate properties of collagen gels[J]. Macromol Res,2019,27(11):1124−1135. doi: 10.1007/s13233-019-7160-y [29] CHEN X, LIU L, YUAN D, et al. Preparation of nano-sized Bi2Te3 thermoelectric material powders by cryogenic grinding[J]. Progress in Natural Science Materials International,2012,3:201−206. [30] 赵绪龙, 马广鹏, 武继锋, 等. 冷冻研磨结合LC-MS/MS法检测指甲中甲基苯丙胺、苯丙胺成分[J]. 刑事技术,2020,6:601−605. [ZHAO X L, MA G P, WU J F, et al. Determination of methamphetamine and amphetamine in nail through frozen grinding coupled into high performance liquid chromatography-tandem mass spectrometry[J]. Forensic Science and Technology,2020,6:601−605. doi: 10.16467/j.1008-3650.2020.06.011ZHAO X L, MA G P, WU J F, et al. Determination of methamphetamine and amphetamine in nail through frozen grinding coupled into high performance liquid chromatography-tandem mass spectrometry[J]. Forensic Science and Technology, 2020, 6: 601-605. doi: 10.16467/j.1008-3650.2020.06.011 [31] BARANOWSKA-KOREZYE A, BARANOWSKA-KOREZYE A, HUDECKI A, et al. Silk powder from cocoons and woven fabric as a potential bio-modifier[J]. Materials,2021,14(22):6919. doi: 10.3390/ma14226919 [32] HESS S, BEEK J V, PANNELLL K. Acid hydrolysis of silk fibroins and determination of the enrichment of isotopically labeled amino acids using precolumn derivatization and high-performance liquid chromatography-electrospray ionization-mass spectrometry[J]. Analytical Biochemistry,2002,311(1):19−26. doi: 10.1016/S0003-2697(02)00402-5 [33] BATH J D, ELLIS J W. Some features and implications of the near infrared absorption spectra of various proteins: Gelatin, silk fibroin, and zinc insulinate[J]. J Phys Chem,2002,45(2):204−209. [34] FRENDEL A, SCHMIDT-NAAKE G. Controlled molecular weight degradation by vibration grinding[J]. 2001, 24(8): 798-803. [35] WANG Y Q, ZHAO P, LIU H, et al. Structure and temperature induced crystallization of natural rubber with different milling times[J]. Polym Sci Ser A,2021,63(3):228−237. doi: 10.1134/S0965545X21030135 [36] AMOURA H, MOKRANE H, NADIEMI B. Effect of wet and dry milling on the functional properties of whole sorghum grain flour and kalian[J]. J Food Sci Technol,2020,57:1100−1109. doi: 10.1007/s13197-019-04145-2 [37] ZHONG J, MA M J, LI W Y, et al. Self-assembly of regenerated silk fibroin from random coil nanostructures to antiparallel β-sheet nanostructures[J]. Biopolymers: Original Research on Biomolecules and Biomolecular Assemblies,2014,101(12):1181−1192. [38] SHAMSHINA J L, STEIN R S, ABIDI N. Choosing the right strategy: Cryogrinding vs. ball milling-comparing apples to apples[J]. Green Chemistry,2021,23(23):9646−9657. doi: 10.1039/D1GC03128G [39] 丁梦瑶, 戴梦男, 李蒙, 等. 不同分子质量丝素蛋白的分离与表征[J]. 纺织学报,2021,42(7):46−53. [DING M Y, DAI M G, LI M, et al. Separation and characterization of silk fibroin with different molecular weight[J]. Journal of Textile Research,2021,42(7):46−53. doi: 10.13475/j.fzxb.20201006308DING M Y, DAI M G, LI M, et al. Separation and characterization of silk fibroin with different molecular weight[J]. Journal of Textile Research, 2021, 42(7): 46-53. doi: 10.13475/j.fzxb.20201006308 [40] ZHOU J, ZHENG D, ZHANG F, et al. A novel method to bind soybean protein onto the surface of poly (Ethylene terephthalate) fabric[J]. Textile Research Journal,2016,87(4):460−473. -
下载:
点击查看大图
图(8)
计量
- 文章访问数: 27
- HTML全文浏览量: 20
- PDF下载量: 4
- 被引次数: 0
