Abd El-Salam Ayat E, Abd-El-Haleem Desouky, Youssef Amany S, Zaki Sahar, Abu-Elreesh Gadallah, El-Assar Samy A
Botany and Microbiology Department, Faculty of Science, Alexandria University, Alexandria, Egypt.
Environmental Biotechnology Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, 21934 Burgelarab, Alexandria, Egypt.
J Genet Eng Biotechnol. 2017 Dec;15(2):335-344. doi: 10.1016/j.jgeb.2017.07.002. Epub 2017 Jul 26.
Among others, isolate PSK1 was selected and identified by 16 S rDNA sequencing as . Growth optimization of PSK1 and physicochemical parameters affected bioflocculant production was carried out by Plackett-Burman design and resulted in increasing in the activity by 4.5%. Bioflocculant production by entrapped and adsorbed immobilized microbial cells was performed using different techniques and revealed enhancement in the activity in particular with pumice adsorption. HPLC analysis of sugars and amino acids composition, FTIR and the effect of different factors on the purified PSK1 biopolymer such as presence of cations, thermal stability, pH range and clay concentration was carried out. Scanning electron microscopy (SEM) of free, immobilized cells, PSK1 bioflocculant and formed flocs were performed. The results revealed that bioflocculant PSK1 is mainly glycoprotein consists of glucose and rhamnose with a large number of amino acids in which arginine and phenylalanine were the major. SEM analysis demonstrated that PSK1 have a clear crystalline rod shaped structure. FTIR spectrum reported the presence of hydroxyl and amino groups which are preferred in flocculation process. PSK1 was soluble in water and insoluble in all other tested organic solvents, while it was thermally stable from 40 to 80 °C. Among examined cations, CaCl was the best coagulant. The maximum flocculation activity of the PSK1 recorded at 50 °C (92.8%), pH 2.0 (94.56%) with clay concentration range 5-9 g/l. To obtain a large amount of PSK1 bioflocculant with high flocculating activity, batch fermentation was employed. The results recorded ∼6 g/l yield after 24 h of fermentation.
其中,分离出的PSK1通过16S rDNA测序进行了选择和鉴定。采用Plackett-Burman设计对PSK1的生长优化以及影响生物絮凝剂产生的理化参数进行了研究,结果使活性提高了4.5%。使用不同技术进行了包埋和吸附固定化微生物细胞产生生物絮凝剂的实验,结果表明特别是浮石吸附可提高活性。对糖和氨基酸组成进行了HPLC分析,对纯化的PSK1生物聚合物进行了FTIR分析,并研究了不同因素如阳离子的存在、热稳定性、pH范围和粘土浓度的影响。对游离细胞、固定化细胞、PSK1生物絮凝剂和形成的絮凝物进行了扫描电子显微镜(SEM)分析。结果表明,生物絮凝剂PSK1主要是糖蛋白,由葡萄糖和鼠李糖以及大量氨基酸组成,其中精氨酸和苯丙氨酸是主要成分。SEM分析表明,PSK1具有清晰的晶体棒状结构。FTIR光谱显示存在在絮凝过程中起作用的羟基和氨基。PSK1可溶于水,不溶于所有其他测试的有机溶剂,在40至80°C时具有热稳定性。在所检测的阳离子中,CaCl是最佳的凝聚剂。PSK1的最大絮凝活性在50°C(92.8%)、pH 2.0(94.56%)、粘土浓度范围为5-9 g/l时记录。为了获得大量具有高絮凝活性的PSK1生物絮凝剂,采用了分批发酵。发酵24小时后,产量记录为约6 g/l。