Isolation and characterization of detergent-compatible amylase-, protease-, lipase-, and cellulase-producing bacteria.

Detergent-compatible enzymes are the new trend followed by most in the detergent industry. Cellulases, lipases, proteases, and amylases are among the enzymes frequently used in detergents. Detergent-compatible enzymes can be obtained from many organisms, but the stability, cheapness, and availability of microbial enzymes make them preferable in industrial areas. In the present study, soil samples contaminated with household waste were collected from different regions of Trabzon (Turkey) for amylase-, cellulase-, protease-, and lipase-producing bacteria. A total of 55 bacterial isolates differing in colony morphology were purified from the samples and 25 of the isolates gave positive results in enzyme screening. The enzyme screening experiments revealed that 10 isolates produced amylase, 9 produced lipase, 7 produced cellulase, and 6 produced protease. While 2 isolates showed both protease and lipase activity, for 2 different isolates cellulose and amylase activity were detected together. It was also observed that one isolate, C37PLCA, produced all four enzymes. The morphological, physiological, and biochemical analyses of the bacteria from which we obtained the enzymes were performed and species close to them were determined using 16S rRNA sequences. Based on the results obtained, our enzymes show tremendous promise for the detergent industry.

Janthinobacterium kumbetense sp. nov., a violacein-producing bacterium isolated from spring water in Turkey, and investigation of antimicrobial activity of violacein.

Strain GKT was isolated from the Kumbet plateu of Giresun in Turkey. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain GKT belonged to genus Janthinobacterium and 16S rRNA gene sequence similarities with all type strains of the genus Janthinobacterium were 98.89%-99.78%. The calculated pairwise average nucleotide identity (ANI) values between strain GKT and all type strains of Janthinobacterium species were in the range of 79.8%-93.2%. In addition, digital DNA-DNA hybridization (dDDH) values were in the range of 23.0%-51.7%. Major fatty acids are C10:03OH, C12:0, C16:1ω7c, C16:0, and C18:1ω7c, and polar lipids included phosphatidylethanolamine, phosphatidylglycerol, also one unidentified phospholipid and one unidentified aminophospholipid. The respiratory quinone of strain GKT was determinated to be Q-8. The genome sizes of strain GKT was 6 197 538 bp with 63.16% G + C ratio. Strain GKT is Gram-stain-negative, aerobic, rod-shaped, and motile. A violet pigment was produced by strain GKT. The crude violacein pigments were separated into three diferent bands on a TLC sheet. Then violacein and deoxyviolacein were purifed by vacuum liquid column chromatography and identifed by NMR spectroscopy. The antimicrobial activities of purifed violacein and deoxyviolacein were screened for seven microorganisms. Based on the results of the morphological, biochemical, physiological, phylogenetic, and genomic characteristics, we propose classifying the strain GKT as representative of a novel species of the genus Janthinobacterium, for which the name Janthinobacterium kumbetense sp. nov. is proposed (GKT = LMG 32662T = DSM 11423T).

Investigation of potential inhibitor properties of violacein against HIV-1 RT and CoV-2 Spike RBD:ACE-2.

A violacein-producing bacterium was isolated from a mud sample collected near a hot spring on Kümbet Plateau in Giresun Province and named the GK strain. According to the phylogenetic tree constructed using 16S rRNA gene sequence analysis, the GK strain was identified and named Janthinobacterium sp. GK. The crude violacein pigments were separated into three different bands on a TLC sheet. Then violacein and deoxyviolacein were purified by vacuum liquid column chromatography and identified by NMR spectroscopy. According to the inhibition studies, the HIV-1 RT inhibition rate of 1 mM violacein from the GK strain was 94.28% and the CoV-2 spike RBD:ACE2 inhibition rate of 2 mM violacein was 53%. In silico studies were conducted to investigate the possible interactions between violacein and deoxyviolacein and three reference molecules with the target proteins: angiotensin-converting enzyme 2 (ACE2), HIV-1 reverse transcriptase, and SARS-CoV-2 spike receptor binding domain. Ligand violacein binds strongly to the receptor ACE2, HIV-1 reverse transcriptase, and SARS-CoV-2 spike receptor binding domain with a binding energy of -9.94 kcal/mol, -9.32 kcal/mol, and -8.27 kcal/mol, respectively. Deoxyviolacein strongly binds to the ACE2, HIV-1 reverse transcriptase, and SARS-CoV-2 spike receptor binding domain with a binding energy of -10.38 kcal/mol, -9.50 kcal/mol, and -8.06 kcal/mol, respectively. According to these data, violacein and deoxyviolacein bind to all the receptors quite effectively. SARS-CoV-2 spike protein and HIV-1-RT inhibition studies with violacein and deoxyviolacein were performed for the first time in the literature.

Change in the Location of the Optic Strut Relative to the Anterior Clinoid Process Pneumatization.

OBJECTIVE: This study aimed to peruse the alteration of the position of the optic strut (OS) according to the anterior clinoid process (ACP) pneumatization. METHODS: This retrospective study conducted on cone-beam computed tomography images of 400 patients with a mean age of 36.49±15.91 years. RESULTS: Anterior clinoid process length, width, and angle were measured as 10.56±2.42 mm, 5.46±1.31 mm, and 42.56±14.68 degrees, respectively. The tip of ACP was measured as 6.60±1.50 mm away from the posterior rim of OS. In the 631 sides (78.87%) did not have ACP pneumatization. In the cases with ACP pneumatization, three different configurations were identified as follows: Type 1 in 71 sides (8.87%), Type 2 in 56 sides (7%), and Type 3 in 42 sides (5.23%). Relative to ACP, the location of OS was determined as follows: Type A in 29 sides (3.64%), Type B in 105 sides (13.12%), Type C in 344 sides (43%), Type D in 289 sides (36.12%), and Type E in 33 sides (4.12%). The spread of data related to the attachment site of OS according to the presence or absence of ACP pneumatization showed that the location of OS was affected by ACP pneumatization ( P <0.001). In ACPs with pneumatization, the frequency of OS position relative to ACP was found as follows: Type A in none of sides (0%), Type B in 8 sides (7.6%), Type C in 53 sides (15.4%), Type D in 88 sides (30.4%), and Type E in 20 sides (60.6%). CONCLUSIONS: The main finding of this study was that the location of OS relative to ACP was affected by ACP pneumatization. In ACPs with pneumatization, OS was located more posteriorly compared with ACPs without pneumatization.

A novel cryptic and theta type plasmid (pHIG22) from Thermus scodotuctus sp. K6.

A cryptic plasmid pHIG22 from Thermus scotoductus sp. K6, an isolate from the Alangullu Hot Spring (Aydin, Turkey), was sequenced and characterized. The pHIG22 plasmid is a multicopy, double stranded and 2222 bp circular molecule with 62.78% GC content, which shows a characteristical nucleotide sequence without any homology to other known plasmids. Five open reading frames were predicted based on the nucleotide sequence analysis. The deduced amino acid sequence of all predicted ORFs didn't show any similarity with any known proteins. Three palindroms were detected and two promoter sequences were predicted in both strands. With electron microscopy (TEM) analysis, the replication intermediates were seen as typical Q-shaped molecules that committing pHIG22 replicates via the Theta replication mechanism. A 2012 bp region among 387 and 614 bp of pHIG22 was determined as minimal replicon which carries the elements necessary for plasmid replication and ori region. Furthermore, quantitative real-time PCR showed that the relative copy number of pHIG22 was estimated to be 148.2 ±â€¯4.7 copies per chromosome equivalent. The new Theta type plasmid would be useful and beneficial to build vectors for cloning of thermophilic genes and in vivo protein engineering.

Evaluation of TNF-alpha gene (G308A) and MBL2 gene codon 54 polymorphisms in Turkish patients with tuberculosis.

OBJECTIVE: MBL acts as a binding protein that enables uptake of mycobacteria into macrophages. And, TNF-alpha is an important cytokine that is involved in control of mycobacterial infections both in-vivo and in-vitro. A large number of genetic factors exerting susceptibility to tuberculosis has been identified, among which mannose-binding lectin and tumor necrosis factor-alpha call attention. The objective of this study is to compare the frequency of TNF-alpha and MBL gene polymorphisms between patients diagnosed with tuberculosis and healthy volunteers in Turkey, and determine the association between tuberculosis and TNF-alpha gene (G308A) and MBL2 gene codon 54 polymorphisms. MATERIAL AND METHODS: The study included 69 patients who were diagnosed with tuberculosis and 70 control subjects. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to detect TNF-alpha (G308A) gene and MBL2 gene codon 54 polymorphisms. For statistical analysis, the significance level was determined as p<0.05. RESULTS: A comparison between patient and control groups in TNF-alpha (G308A) gene and MBL2 gene codon 54 polymorphisms showed no statistically significant difference (p>0.05). However, a comparison of mean body mass index (BMI) and smoking status showed a statistically significant difference between the tuberculosis and control groups (p=0.01 and p=0.009, respectively). CONCLUSION: Our results suggest that the MBL2 gene Codon 54 and TNF-alpha gene G308A polymorphisms are not associated with an increased risk for development of tuberculosis in our patients. Further studies are required including more cases of tuberculosis patients and other potentially relevant gene polymorphisms.

Relief from sleep apnea after radiation and chemotherapy.

Superior vena cava syndrome (SVCS) can result from extrinsic compression by a primary tumor, mediastinal lymph nodes metastases, benign lesions, or intraluminal thrombosis. The association between obstructive sleep apnea and SVCS has not been extensively evaluated. To our knowledge, only 5 cases of obstructive sleep apnea in SVCS have been reported in the literature. We presented a 53-year-old man who was admitted with dyspnea, edema of the face, and excessive daytime sleepiness. Chest radiography and computed tomography revealed lung cancer. A biopsy of the tumor revealed squamous cell carcinoma. Obstructive sleep apnea was diagnosed by polysomnography (apnea hypopnea index: 13 per hour). After radiation and chemotherapy, edema of the face, snoring, and daytime sleepiness were alleviated, and the patient's apnea hypopnea index decreased to 0.6 per hour. In conclusion, there is a relationship between obstructive sleep apnea and SVCS.