T mostly of two different varieties of -chains (1 and 2) and dimeric -chains. The molecular weight of collagen was analyzed using Quantity A single four.six.0 application (Bio-Rad Laboratories, Hercules, CA, USA); we located that the molecular weight of ASC (1-MW, 137 kDa; 2-MW, 127 kDa) was slightly larger than that of PSC (1-MW, 135 kDa; 2-MW, 123 kDa), which might be attributed for the removal of telopeptide regions from the PSC . The protein patterns of ASC and PSC had been equivalent to these of your collagen obtained from tilapia skin  and Pacific cod skin . While Etiocholanolone medchemexpress pepsin removed the cross-link-containing telopeptide, the electrophoresis patterns showed that PSC contained a larger intensity of -chains than ASC, indicating that PSC has high molecular cross-linkages [23,24]. In addition, the ratio of 1 and 2 was calculated by Image J software (VERSION 1.eight.0, National Institute of Mental Overall health, Bethesda, MD, USA); GS-626510 Purity specifically, the ratios of 1 and two for ASC and PSC have been 1.86 and 2.23, respectively, both close to two:1, implying that ASC and PSC extracted from lizardfish scales are form I collagen (2 two) .Mar. Drugs 2021, 19, 597 Mar. Drugs 2021, 19, x FOR PEER Assessment Mar. Drugs 2021, 19, x FOR PEER REVIEW3 of 17 three of 18 4 of2.3.three. Circular Dichroism (CD) Spectrum CD can be a very simple and powerful approach to identify no matter if the triple helical structure is intact . The CD spectrum of native collagen using a triplehelix structure shows a constructive peak at 221 nm (maximum positive cotton effect), a damaging peak at 198 nm (max imum adverse cotton effect), along with a crossover point (zero rotation) at approximately 213 nm [10,22]. As shown in Figure 2c, the CD spectrum of lizardfish scales ASC and PSC exhibited weak good absorption peaks at 221 nm and 220 nm, respectively, and nega tive absorption peaks have been observed at 198 nm and 197 nm, respectively, both with a crossover point at 213 nm. Furthermore, the Rpn values (the ratio from the good to adverse) of ASC and PSC have been 0.12 and 0.14, respectively, indicating that the collagen extracted from lizardfish scales possess a triplehelix conformation [26,27].Figure 1. SDSPAGE patterns of ASC and PSC from lizardfish scales. Lane 1: Marker regular; Lane Figure 1. SDS-PAGE patterns of ASC and PSC from lizardfish scales. Lane 1: Marker regular; two: PSC; Lane 3: ASC. The experiment was performed only when (n = 1). Lane 2: PSC; Lane 3: ASC. The experiment was conducted only once (n = 1). 2.3.four. Xray Diffraction (XRD) Spectrum2.three.The XRD patterns of ASC and PSC are shown in Figure 2d. We found that ASC and Spectroscopy Characterization two.three. Spectroscopy Characterization PSC consisted of two peaks, a sharp and also a broad peak. The diffraction angles (2) of ASC 2.3.1. UV Absorption Spectrum 2.three.1. UV Absorption Spectrum have been 7.86and 21.25 and these of PSC had been 7.58and 21.02 which are consistent with Normally, collagen has a maximum absorption peak in 21040 nm range, that is Typically, collagen has a maximum absorption peak in 21040 nm range, which can be the characteristic diffraction peaks of collagen . The d worth from the very first sharp peak of attributed for the presence of C=O, OOH, and CONH2 2groups in the polypeptide chains attributed towards the presence of C=O, OOH, and CONH groups in the polypeptide chains ASC was 11.25 and that of PSC was 11.66 and this reflects the distance amongst the of collagen . The UV absorption spectra of lizardfish scales coll.