Sys Rev Pharm 2020;11(7):06-09 A multifaceted review journal in the field of pharmacy 6 Systematic Reviews in Pharmacy Vol 11, Issue 7, July-Aug 2020 Investigation of Crystallinity Characterization of Bamboo Fibers Using Xylanase from Aspergillus nidulans Noor T. Hamdan Biology Department, College of Science, Mustansiriyah University, Baghdad- Iraq. Corresponding author email: noor.t.hamdan@uomustansiriyah.edu.iq ABSTRACT The Crystallinity phenomenon refers to a crucial feature of lignocellulosic biomass due to its substantial impact on enzyme treatment. The X-ray diffraction analysis is applied to determine the parameters characteristic of the bamboo fibers such as crystallinity index, crystalline %, crystallite size as well as Z- value. Results of the X-ray diffraction analysis appeared maximum in treated bamboo fiber at temperature 40 ºC. They estimated the highest value in 57% of crystallinity index, 72% of crystalline %, 3.91nm of crystallite size and -34.6 of Z- value in treated bamboo fiber than untreated bamboo fiber. This increase could be attributed to the completely removal of amorphous non-cellulosic compounds by xylanase treatment, which would give better smoothing of the surface of bamboo fiber. Keywords: Xylanase, crystallinity index, crystallite size, crystalline %, Bamboo fiber, Z- value. Correspondence: Noor T. Hamdan Biology Department, College of Science, Mustansiriyah University, Baghdad- Iraq. Corresponding author email: noor.t.hamdan@uomustansiriyah.edu.iq INTRODUCTION Cellulose refers to a polymer of β-glucose with β-1-4 bonds between the units of glucose. There are two regions within cellulose. The crystalline region of cellulose constituted of highly ordered cellulose molecules reaching 2/3 of the whole cellulose, whereas within in the amorphous region contains less ordered molecules [1]. These crystalline regions and amorphous regions have been identified by X-ray diffraction analysis such as crystallinity index, crystalline %, crystallite size as well as Z- value. Bamboo fiber termed as one of the lignocelluloses producing plants which contains cellulose 73.8%, hemicellulose 12.5%, lignin 10.1%, and pectin 0.4% [2]. Bamboo fiber has some major disorders such as harsh prickly feel and poor drapeabity. Therefore, it is necessary to remove these disorders through biological processing by biotechnology termed xylanase treatment [3]. The enzymatic approach of fibers consider an environmental process due to enhancing of the characteristic of the fiber, their desirable surface as well as the soft handle [4]. Our study aimed to determine the characteristics of the X- ray diffraction of bamboo fibers using xylanase from Aspergillus nidulans. Materials and Methjods Enzyme treatment of the bamboo fibers The material used in this work was raw bamboo fiber which was obtained from agricultural nursey in Iraq. The bamboo clum was chopped into small chips after removing the epidermis from bamboo and then chopped into small chips. After that their small chips were put in a hammer mill and then grounded, sieved using a 100µm sieve. The bamboo powder dried 72h in an oven at 70ºC and placed in a glass jar for further analysis. Dried bamboo powders weighing 200mg were immersed in test tubes containing 10ml (50mM, pH7.2) phosphate buffer and 10ml of 100% crude xylanase enzyme dosage. The tubes were incubated for six hours at 30°C, 40°C and 50°C. Test tubes were then boiled in a water bath for 5 min. to finish enzyme activity and cooled at room temperature. After that, the reducing sugars were determined in the incubation mixture as described in Ghose,(1987) [5]. After that, these bamboo fibers were air-dried and used as further study. X-Ray Diffraction (XRD) studies The structural characteristic of the raw bamboo fibers, as well as treated bamboo fibers, were achieved to measure the crystallinity degree by wide-angle of X-ray diffractogram. The powder fibers were examined between angles (2θ) 5° to 70° to obtain the equatorial reflections using X-ray diffractometer (XRD) Bruker AXS D8 with CuKα radiation (λ- 1.5418Å). The generator was utilized at 40KV, 30mA at a scanning level of 1.2°/ 1min. The crystallinity index (C.I) was determined by the equation as follows (Sajithkumar et al., 2016) [6]: C.I% = I 200− I am I 200 × 100 Where I 200 the intensity of the peak from 200 lattice plane (2θ=22.4º) representing crystalline material while I am the diffracted intensity peak at 2θ =18º indicating amorphous material of bamboo fibers. The percent crystallinity of the raw and treated bamboo fibers of each temperature 30 ºC, 40 ºC, and 50 ºC was evaluated using the equation given below (Kaith and Kalia, 2008) [7]: Crystalline % = I 22 I 22 + I 18 × 100 Where I 22 and I 18 are the 2θ scale of 22º and 18º representing the crystalline and amorphous intensities respectively. The crystallite size of the fibers samples was evaluated using the following Scherrer formula (Han et al., 2014) [8]: L = Kλ / β Cos θ Where L, the crystallite size; K, the shape factor (0.94); λ, the X-ray wavelength used; B, the half-width maximum of the equatorial reflections and θ, Bragg angle related to the 200 plane. The Z-discriminant (Z-value) analysis was achieved for the differentiation of cellulose whether Iα (monoclinic) or Iβ (triclinic) of the crystalline cellulose structure in bamboo fibers was utilized (Wada and Okano., 2001) [9]. This analysis is determined by Equation (Poletto et al., 2012) [10]: Z = 1693d1 – 902d2 – 594