Muslim Sahira Nsayef, Al-Kadmy Israa M S, Auda Ibtesam Ghadban, Mohammed Ali Alaa Naseer, Al-Jubori Sawsan Sajid
Mustansiryiah University, College of Science, Department of Biology, Branch of Biotechnology, Baghdad, Iraq POX 10244.
J AOAC Int. 2018 Sep 1;101(5):1623-1630. doi: 10.5740/jaoacint.17-0422. Epub 2018 Apr 4.
Lectin was initially called hemagglutinin or agglutinin because of its capacity to agglutinate human as well as human erythrocytes. They are a heterogeneous group of proteins or glycoproteins of nonimmune origin. Because of their chemical properties, they have become a useful tool in several fields such as immunology, cell biology, molecular biology, membrane structure, pharmacology, cancer research, clinical chemistry, and genetic engineering.
The wide applications of lectins users urged the need to isolate lectins from a new strain of bacteria can produce new and high yield of lectin because the current production of lectin from Pseudomonas spp. is very expensive. The goal of this study was to screen the ability of Acinetobacter baumannii isolates to produce lectin and detection of its phenotypic and genotypic profiles and detection of lectin ability to inhibit ofbiofilm formation.
Fifty-one isolates from different sources were collected and detected genetically by using the recA gene. Phenotypic detection of lectin by using semi-quantitative analysis and quantitative analysis in microtiter plate. Genotypic detection of lectin by designed lec gene and used PCR technique. The lectin was extracted by using glass beads and purified by chromatographyic technique followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for determination the molecular size of lectin and finally detection the spectrum of biofilm inhibition by the purified lectin toward biofilm producers.
Of 51 A. baumannii isolates, 17 (33.3%) have been found to produce lectin. Ten of 17 were sequenced, of which 2 were submitted and tested by the gene bank National Center for Biotechnology Information (NCBI), and accession numbers (KX766405.1 and KX766406.1) were obtained. These 17 isolates were phenotypically and genotypically positive for lectin and showed different lec gene expression in semi-quantitative and quantitative analysis. The activities ranged between 4-128 U/mL. Lectin purified by ammonium sulfate precipitation was used to inhibit biofilm formation. We found reduction at three different types of bacteria ranging from 26% for Klebsiella pneumonia, 46.7% for P. stutzeri and 53% for A. baumannii. These results suggested that lectin has a promising application as an antibiofilm agent to combat the growing number of multidrug-resistant pathogen-associated infections.
Lectin has been detected recently in A. baumannii, but the genetic property of this lectin has not yet been fully studied. In our study, we determined the presence of the lectin gene (lec gene) in A. baumannii by using PCR technique, and lec PCR products were identified with various source of isolation and sequenced to screening for epidemiology and submitted to the gene bank NCBI under accession number (KX766405.1 and KX766406.1).
A. baumannii has an ability to produce lectin protein; Lec gene was detected in A. baumannii, and the sequence was recorded under accession number KX766405.1 and KX766406.1.; Lectin was extracted by glass beads and purified by chromatographyic technique; Lectin had strong effect against biofilm formation.
凝集素最初被称为血凝素或凝集原，因为它能够凝集人类以及人类红细胞。它们是一组非免疫来源的异质性蛋白质或糖蛋白。由于其化学性质，它们已成为免疫学、细胞生物学、分子生物学、膜结构、药理学、癌症研究、临床化学和基因工程等多个领域的有用工具。
凝集素使用者的广泛应用促使人们需要从一种新的细菌菌株中分离凝集素，因为从假单胞菌属目前生产凝集素非常昂贵，而新菌株可以产生新的高产凝集素。本研究的目的是筛选鲍曼不动杆菌分离株产生凝集素的能力，检测其表型和基因型特征，并检测凝集素抑制生物膜形成的能力。
从不同来源收集51株分离株，使用recA基因进行基因检测。通过在微量滴定板中进行半定量分析和定量分析来进行凝集素的表型检测。通过设计lec基因并使用PCR技术进行凝集素的基因型检测。使用玻璃珠提取凝集素，并通过色谱技术进行纯化，随后进行十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳以确定凝集素的分子大小，最后检测纯化的凝集素对生物膜产生菌的生物膜抑制谱。
在51株鲍曼不动杆菌分离株中，发现17株（33.
