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Neuromyelitis optica spectrum disorders (NMOSDs) are rare autoimmune-mediated chronic inflammatory disorders involving the central nervous system. The concept of NMOSDs has widened up recently. Its diagnostic criteria have broadened the knowledge of this particular disease including the atypical cases where antiaquaporin-4 antibody is negative and anti-myelin oligodendrocyte glycoprotein (MOG) antibody is positive. Myelin oligodendrocyte glycoprotein is a protein expressed on the outer surface of myelin sheath and oligodendrocytes of the central nervous system. The detection of anti-MOG-Ab is emerging evidence, and thus, research on the role of this antibody in such seronegative cases is still underway. Early diagnosis and adequate therapy is essential in such cases. Here, we report an eight-year-old girl who was diagnosed with NMOSDs and found to have anti-MOG antibodies in place of AQP-4 antibodies.

Background/aimsTo evaluate the corneal endothelial changes in patients with alcohol dependence syndrome (ADS).MethodsA total of 322 corneas of 322 subjects were studied, and two groups were formed. The first, the ADS group, included 161 corneas of 161 patients and the second, the age-matched control group, was composed of 161 corneas of 161 healthy subjects. Both the groups were examined by specular microscope and the endothelial parameters were compared. The endothelial parameters were also compared before and after the abstinence of alcohol in the former group. The ADS group was further analysed to assess the influence of life time alcohol consumption, years/pattern of drinking, levels of Liver enzymes, levels of mean corpuscular volume, screening questionnaires and stages of fatty liver on corneal endothelium.ResultsOn comparing the two groups using Mann-Whitney U test, a significant difference was found in central corneal thickness (CCT) and endothelial cell density (CD) between the ADS (CCT: 529±29 µm, CD: 2571±236 cells/mm2) and control groups (CCT: 510±32 µm, CD: 2752±291 cells/mm2) (PConclusionsCorneal endothelial alteration is present in patients with ADS, more marked in patients with an increase in stage of fatty liver.

This study investigated the effect of dietary supplemented synthetic, and shrimp shell meal (SSM) derived natural astaxanthin combination on growth, digestive and metabolic enzyme activities, antioxidative status, physio-metabolic responses, and skin colouration of high valued discus (Symphysodon aequifasciatus) advanced juveniles reared in captive condition. Four isonitrogenous (50 % crude protein), isolipidic (9% ether extract), and isocaloric (around 400 kcal digestible energy/100 g) experimental diets viz. Control (only synthetic astaxanthin, without SSM), T1 (15 % SSM + synthetic astaxanthin), T2 (20 % SSM + synthetic astaxanthin), T3 (25 % SSM + synthetic astaxanthin) were prepared and fed to satiation level twice daily for 90 days. Results indicated significantly (P

Carotenoprotein, a complex of carotenoids and protein, extracted from shrimp shell waste can be an effective source of carotenoid as well as antioxidant in fish diets. In the present study, the nutritional potential of carotenoprotein extracted from shell wastes of Parapeneopsis stylifera on growth, physiometabolic changes and coloration in high-value ornamental fish, oscar (Astronotus ocellatus) was assessed in a 120 days feeding trial. Four iso‑nitrogenous (30–31% CP) and isolipidic (11.34–11.55% CL) diets were formulated with graded levels of carotenoprotein inclusion like 0%, 5%, 10% and 15% and designated as Control, T1, T2 and T3 respectively. At the end of the feeding trial, growth indices such as final body weight, weight gain, and specific growth rate indicated that fish fed with 10 and 15% levels of carotenoprotein incorporated diet showed higher growth compared to control and T1. Oxidative stress enzymes like SOD and Catalase were lower in 15% and 10% carotenoprotein incorporated diets respectively in both gills and liver. Stress indicators like serum glucose and cortisol activity were found to be higher in control than in diets with 10% (T2) as well as 15% (T3), suggesting the stress-reducing ability of the carotenoprotein in fish feed. Relatively higher respiratory burst activity in fishes of T3 group indicated higher immunity status of the fish. Quantitative colour values in terms of redness, chroma in the fin region as well as carotenoid content in scale, skin and muscle of fish were higher in T2 (10% carotenoprotein incorporated diet) as compared to T3 (15% carotenoprotein incorporated diet). Overall, this study clearly showed that dietary incorporation of as little as 10% carotenoprotein is advisable to ornamental fish farmers for enhanced growth, colouration, as well as antioxidant activity in this popular high value ornamental fish for maximizing their profit.

PURPOSE To evaluate the corneal endothelial changes in patients with chronic renal failure. METHODS A total of 128 corneas of 128 subjects were studied, and 3 groups were formed. The first, the dialyzed group, composed of 32 corneas of 32 patients; the second, the nondialyzed group, composed of 34 corneas of 34 patients; and the third, the age-matched control group, composed of 64 corneas of 64 healthy subjects were examined by a specular microscope and the endothelial parameters were compared. The dialyzed group (enhanced level of toxins in the blood) was further analyzed to assess the influence of blood urea, serum creatinine, serum calcium, and serum phosphorus including the duration of dialysis on corneal endothelium. RESULTS On comparing the 3 groups using analysis of variance and posthoc tests, a significant difference was found in the central corneal thickness (CCT) and endothelial cell density (CD) between the control (CCT: 506 ± 29 μm, CD: 2760 ± 304 cells/mm) and dialyzed groups (CCT: 549 ± 30 μm, CD: 2337 ± 324 cells/mm) [P < 0.001 (CCT); P < 0.001 (CD)]; control and nondialyzed groups (CCT: 524 ± 27 μm, CD: 2574 ± 260 cells/mm) [P = 0.023 (CCT); P = 0.016 (CD)]; and dialyzed and nondialyzed groups [P = 0.002 (CCT); P = 0.007 (CD)]. Using the linear generalized model, a significant correlation was found between the endothelial parameters and blood urea only [P = 0.006 (CCT), 0.002 (coefficient of variation), 0.022 (CD), and 0.026 (percentage of hexagonality)], although the correlation was poorly positive for CCT but poorly negative for the remaining endothelial parameters. CONCLUSIONS Corneal endothelial alteration is present in patients with chronic renal failure, more marked in patients undergoing hemodialysis and with raised blood urea level.

Introduction: India is presently facing an epidemic of diabetes mellitus and the risks of chronic complications from the disease are associated with the duration of the disease as well as the degree of hyperglycaemia. Diabetic retinopathy is a known microvascular complication of diabetes mellitus and is the most common cause of blindness in the western countries. Apolipoproteins are the protein component of lipoproteins. Apart from acting as structural proteins, they also act as cofactors to various enzymes. Aim: To measure the levels of serum apolipoproteins and glycated haemoglobin in cases of diabetic retinopathy and to assess their association with the stages of diabetic retinopathy. Material and Methods: The 135 diabetic cases [with (110) and without (125) retinopathy] attending the Ophthalmology OPD of this tertiary care hospital were included in the present study. Following retinoscopy, the patients were classified as Non-Proliferative Diabetic Retinopathy (NPDR) (n=75) and Proliferative Diabetic Retinopathy (PDR) (n=35). The controls (n=100) were age and sex matched patients who did not have diabetes. The cases and controls were assessed for HbA1c, total cholesterol, triglycerides, HDL cholesterol, Apo A-I and Apo B-100. Results: The HbA1c was found to be higher in diabetics without retinopathy (7.02%) as compared to controls (5.58%) (p

Carotenoproteins from four different shrimp shell wastes Penaeus monodon, Parapenaeopsis stylifera, Metapenaeus affinis and Nematopalemon tenuipes were extracted with the aid of papain enzyme and characterized by their protein, amino acid and carotenoid content of the shell wastes and the antioxidant activities like DPPH, FRAP, ABTS radical scavenging activity and reducing power assay of the carotenoprotein. Higher protein content of 9.8 g 100 g−1 and 9.2 g 100 g−1 was recovered from shell waste of Penaeus monodon and Parapenaeopsis stylifera respectively along with highest carotenoid content of 114 ± 0.02 μg g−1 in Parapenaeopsis stylifera followed by 100.6 ± 0.02 μg g−1 from the shell waste of Metapenaeus affinis. Highest antioxidant activity was found in the carotenoprotein extracted from the shell waste of P. stylifera which suggest that the antioxidant activity of carotenoids followed a concentration dependent pattern. The amino acid profile showed that carotenoprotein is a rich source of essential amino acids such as glutamic acid, aspartic acid, lysine and leucine. Among shell wastes, P.stylifera shell waste was calculated to be superior as it contained higher amount of essential amino acids and exhibited higher antioxidant activity in terms of protein, carotenoid as well as radical scavenging and reducing power and it could serve as a supplementary nutritive feed ingredient in animal diets. This would help in utilization of crustacean (shrimp) shell waste for formulating low cost feed for ornamental fish and also encourage shrimp processing industries to utilize of the same in order to control pollution of land and water.

Background: Optical coherence tomography is a non-invasive imaging technique routinely used in ophthalmology to visualize and quantify the layers of the retina. It also provides information on optic nerve head topography, peripapillary retinal nerve fibre layer thickness and macular volume which correlate with axonal loss. These measurements are of interest in optic neuropathies and in multiple sclerosis. The OCT parameters are now used as endpoints in neurologic clinical trials.Methods: A prospective study involving 30 patients of multiple sclerosis and equal number of age and sex matched controls were subjected to evaluation of retinal changes (peripapillary retinal nerve fiber layer and central macular thickness) using Zeiss Cirrus HD-OCT machine. The results collected were then subjected to statistical analysis.Results: Significant RNFL thinning was seen in patients of multiple sclerosis compared to the age and sex matched controls. Marked thinning was seen in superior and temporal quadrants of right eye (p value of 0.002 and 0.008 respectively) and all quadrants in left eye with a p value of < 0.001. Patients with multiple sclerosis for more than 5 yrs showed statistically significant RNFL thinning in the superior quadrant of right eye (p

The elongation factor Tu (EF-Tu) from Pseudomonas aeruginosa was purified as a 45-kDa polypeptide that forms a complex with both the 12- and 16-kDa forms of nucleoside-diphosphate kinase (Ndk) and predominantly synthesizes GTP. 70 S ribosomes of P. aeruginosa predominantly synthesize GTP, which is inhibited in presence of anti-Ndk antibodies. Anti-EF-Tu antibodies change the specificity of ribosomal GTP synthesis to all nucleoside triphosphate synthesis. Ndk has been shown to be a part of 30 S ribosomes, whereas EF-Tu is found to be associated with the 50 S ribosomal subunit. These data indicate that GTP synthesis in the ribosome is modulated both by Ndk and by EF-Tu. Peptide chain elongation as measured by polymerization of Phe-tRNA on a poly(U) template in presence of GDP can be inhibited by anti-Ndk antibodies and restored by the addition of GTP. Anti-EF-Tu antibodies similarly inhibit peptide chain elongation by P. aeruginosa ribosomes in the in vitro translation assay; however, this inhibition cannot be overcome by adding back GTP. Because the purified EF-Tu.16-kDa Ndk complex predominantly synthesizes GTP, it seems likely that this complex is a significant source of GTP for translational elongation in protein biosynthesis.

We report the purification and characterization of a protein from the membrane fraction of Pseudomonas aeruginosa showing intrinsic guanosine triphosphatase (GTPase) activity. The protein was purified as a 48-kDa polypeptide capable of binding and hydrolyzing GTP. The N-terminal sequence of the purified protein revealed its similarity to the Escherichia coli Ras-like protein (Era), and the protein cross-reacted with anti-Era antibodies. This protein was named Pseudomonas Ras-like protein (Pra). Anti-Pra antibodies also cross-reacted with E. coli Era protein. Pra is autophosphorylated in vitro, with phosphotransfer of the terminal phosphate from [gamma-32P]GTP but not [gamma-32P]ATP. Pra is capable of complex formation with the truncated 12-kDa form of nucleoside diphosphate kinase (Ndk) but not with the 16-kDa form. Purified Pra was also shown to physically interact with pyruvate kinase (Pk); Pk and Pra can form a complex, but when the 12-kDa Ndk, Pk, and Pra are all present, Pk has a higher affinity than Pra for forming a complex with the 12-kDa Ndk. The 12-kDa Ndk-Pra complex catalyzed increased synthesis of GTP and dGTP and diminished synthesis of CTP and UTP or dCTP and dTTP relative to their synthesis by uncomplexed Ndk. Moreover, the complex of Pra with Pk resulted in the specific synthesis of GTP as well when Pra was present in concentrations in excess of that of Pk. Membrane fractions from cells harvested in the mid-log phase demonstrated very little nucleoside triphosphate (NTP)-synthesizing activity and no detectable Ndk. Membranes from cells harvested at late exponential phase showed NTP-synthesizing activity and the physical presence of Ndk but not of Pk or Pra. In contrast, membrane fractions of cells harvested at early to late stationary phase showed predominant GTP synthesis and the presence of increasing amounts of Pk and Pra. It is likely that the association of Pra with Ndk and/or Pk restricts its intrinsic GTPase activity, which may modulate stationary-phase gene expression and the survival of P. aeruginosa by modulating the level of GTP.

We report the utilization of site-directed and random mutagenesis procedures in the gene encoding nucleoside diphosphate kinase (ndk) from Pseudomonas aeruginosa in order to examine the role of Ndk in the production of alginate by this organism. Cellular levels of the 16-kDa form of the Ndk enzyme are greatly reduced in P. aeruginosa 8830 with a knockout mutation in the algR2 gene (8830R2::Cm); this strain is also defective in the production of the exopolysaccharide alginate. In this study, we isolated four mutations in ndk (Ala-14-->Pro [Ala14Pro], Gly21Val, His117Gln, and Ala125Arg) which resulted in the loss of Ndk biochemical activity; hyperexpression of any of these four mutant genes did not restore alginate production to 8830R2::Cm. We identified six additional amino acid residues (Ser-43, Ala-56, Ser-69, Glu-80, Gly-91, and Asp-135) whose alteration resulted in the inability of Ndk to complement alginate production. After hyperproduction in 8830R2::Cm, it was determined that each of these six mutant Ndks was biochemically active. However, in four cases, the in vivo levels of Ndk were reduced, which consequently affected the growth of 8830R2::Cm in the presence of Tween 20. Two mutant Ndk proteins which could not complement the alginate synthesis defect in 8830R2::Cm were not affected in any characteristic examined in the present study. All of the mutant Ndks characterized which were still biochemically active formed membrane complexes with Pk, resulting in GTP synthesis. Two of the four Ndk activity mutants (His117Gln and Ala125Arg) identified were capable of being truncated to 12 kDa and formed a membrane complex with Pk; however, the complexes formed were inactive for GTP synthesis. The other two Ndk activity mutants could be truncated to 12 kDa but were not detected in membrane fractions. These results further our understanding of the role of Ndk in alginate synthesis and identify amino acid residues in Ndk which have not previously been studied as critical to this process.

Nucleoside diphosphate kinase (EC 2.7.4.6) (Ndk) is a ubiquitous enzyme functioning in the intracellular distribution of terminal phosphate bond energy among the various nucleotides used in synthetic and regulatory functions in cells. We have previously reported that in Pseudomonas aeruginosa, this important enzyme is transcriptionally regulated by the gene algR2 and posttranslationally regulated by a phosphoprotein phosphatase for the phosphorylated form of Ndk. We report here that an intracellular protease cleaves the 16-kDa form of Ndk to a 12-kDa form that undergoes autophosphorylation with an efficiency almost identical to that of the 16-kDa form. The 12-kDa form was found to be predominantly associated with the P. aeruginosa cell membrane fraction, whereas the 16-kDa form was predominantly cytoplasmic. In the membrane-associated state, the 12-kDa form of Ndk was found to synthesize GTP in preference to other nucleoside triphosphates. The specificity toward GTP synthesis could be abolished by the addition of Tween 20 or Triton X-100. The activity itself could be abolished by the addition of anti-Ndk antibody to the assay mixture. The formation of the 12-kDa form of Ndk and its association with the cell membrane were found to be related to the growth stage of P. aeruginosa, with less than 1% of the 12-kDa Ndk detectable in the membrane fraction at early log phase in comparison with the levels present at late stationary phase.

Burkholderia cepacia AC1100 uses the chlorinated aromatic compound 2,4,5-trichlorophenoxyacetic acid as a sole source of carbon and energy. The genes encoding the proteins involved in the first step (tftA and tftB [previously designated tftA1 and tftA2, respectively]) have been cloned and sequenced. The oxygenase, TftAB, is capable of converting not only 2,4,5-trichlorophenoxyacetic acid to 2,4,5-trichlorophenol but also a wide range of chlorinated aromatic phenoxyacetates to their corresponding phenolic derivatives, as shown by whole-cell and cell-free assays. The rate of substrate utilization by TftAB depends upon the extent of chlorination of the substrate, the positions of the chlorines, and the phenoxy group. These results indicate a mechanistic similarity between TftAB and the 2,4-dichlorophenoxyacetic acid/alpha-ketoglutarate-dependent dioxygenase, TfdA, from Alcaligenes eutrophus JMP134. The promoter of the oxygenase genes was localized by promoter-probe analysis, and the transcriptional start site was identified by primer extension. The beta-galactosidase activity of the construct containing the promoter region cloned upstream of the beta-galactosidase gene in the promoter-probe vector pKRZ-1 showed that this construct is constitutively expressed in Escherichia coli and in AC1100. The -35 and -10 regions of the oxygenase genes show significant sequence identity to typical Escherichia coli sigma 70 promoters.

Nucleoside-diphosphate kinase (ATP:nucleoside-diphosphate phosphotransferase, EC 2.7.4.6; NDP kinase) is an important enzyme for the maintenance of the correct cellular levels of nucleoside triphosphates (NTPs) and their deoxy derivatives (dNTPs) and is involved in the regulation of cellular development. The enzyme is under the dual control of algR2 and algH in Pseudomonas aeruginosa. We report here the purification and characterization of a protein that dephosphorylates the phosphorylated intermediate form of the P. aeruginosa NDP kinase (Ndk). Dephosphorylation of Ndk phosphate leads to loss of its enzymatic activity. The 10.1-kDa polypeptide shows 77% homology at the N terminus with the Spo0E phosphatase, identified as a negative regulator of sporulation in Bacillus subtilis and 66% with the human Bax protein, identified as an effector of programmed cell death. The phosphatase termed Npp showed varied specificity toward phosphorylated Ndks from different sources including human erythrocyte Ndk phosphate. Its activity toward other histidine phosphates such as CheA or the alpha-subunit of succinyl-CoA synthetase or phosphoesters such as p-nitrophenyl phosphate was quite limited. Npp was stable at room temperature up to 2 h and required Mg2+ for activity. The presence of a phosphatase capable of dephosphorylating the phosphorylated form of P. aeruginosa Ndk represents an interesting and efficient mode of post-translational modification of an enzyme crucial to cellular development.

We have previously reported that two genes cloned from a cosmid library of Escherichia coli can restore mucoidy to an algR2 mutant of Pseudomonas aeruginosa. AlgR2 is a protein involved in the regulation of nucleoside diphosphate kinase (Ndk) as well as alginate synthesis in P. aeruginosa. One of the E. coli genes, rnk, encodes a 14.9 kDa protein with no homology to any other proteins. The other gene, sspA, encodes the stringent starvation protein, a regulatory protein involved in stationary-phase regulation and the stringent response of E. coli. While both rnk and sspA restored alginate production to the P. aeruginosa algR2 mutant, only rnk restored Ndk activity to the mutant. In this report, we have examined the effect of mutations in rnk and sspA on the levels of Ndk in E. coli. We find that a mutation in rnk drastically reduces the level of Ndk in E. coli. A mutation in sspA, however, affects the level of another nucleoside diphosphate kinase distinct from Ndk. The proteins can be easily distinguished from each other by their different affinities for nucleoside diphosphates (NDPs) and also by the differential effect of anti-Ndk antibodies on the reactions they catalyse. The ability of either of these two proteins to restore alginate synthesis in the algR2 mutant of P. aeruginosa demonstrates the importance of nucleoside triphosphate synthesis and energy metabolism for alginate synthesis. Additionally, a role for the stringent starvation protein (SspA) in the modulation of nucleoside triphosphate (NTP) levels in E. coli is also suggested from these experiments.

Alginate is an important virulence factor for Pseudomonas aeruginosa during infection of the lungs of cystic fibrosis patients. The genes encoding enzymes for alginate production by P. aeruginosa are normally silent. They are activated in response to several environmental conditions, including high osmolarity, exposure to ethanol, or long-term growth under conditions of nutrient deprivation. Several genes which participate in the activation of alginate gene promoters have been identified; among these is the algR2 (algQ) gene. AlgR2 is an 18-kDa protein which has been shown to regulate the critical algD gene encoding GDP-mannose dehydrogenase as well as to regulate the levels of a tricarboxylic acid cycle enzyme, i.e., succinyl coenzyme A synthetase, and nucleoside diphosphate kinase (Ndk), an enzyme involved in nucleoside triphosphate synthesis. Succinyl coenzyme A synthetase and Ndk form a complex in P. aeruginosa. While algR2 is required for alginate synthesis at 37 degrees C, an algR2 insertion mutant was still able to make alginate slowly at 37 or at 30 degrees C. We used this observation to identify and clone a gene, termed algH. A strain with mutations in both algR2 and algH is unable to produce alginate at either 37 or 30 degrees C, and it is fully defective in Ndk production.

Infection with mucoid, alginate-producing strains of Pseudomonas aeruginosa is the leading cause of mortality among patients with cystic fibrosis. Alginate production by P. aeruginosa is not constitutive but is triggered by stresses such as starvation. The algR2 (also termed algQ) gene has been previously identified as being necessary for mucoidy; an algR2 mutant strain is unable to produce alginate when grown at 37 degrees C. We show here that the levels of phosphorylated succinyl coenzyme A synthetase (Scs) and nucleoside diphosphate kinase (Ndk), which form a complex in P. aeruginosa, are reduced in the algR2 mutant. We were able to correlate the lower level of phosphorylated Scs with a decrease in Scs activity. Western blots (immunoblots) also showed a decreased level of Ndk in the algR2 mutant, but the presence of another kinase activity sensitive to Tween 20 provides the missing Ndk function. The effect of AlgR2 on tricarboxylic acid (TCA) cycle enzymes appears to be specific for Scs, since none of the other TCA cycle enzymes measured showed a significant decrease in activity. Furthermore, the ability of the algR2 mutant to grow on TCA cycle intermediates, but not glucose, is impaired. These data indicate that AlgR2 is responsible for maintaining proper operation of the TCA cycle and energy metabolism.

The regulatory protein AlgR2 in Pseudomonas aeruginosa positively regulates nucleoside diphosphate kinase (Ndk) and succinyl-CoA synthetase, enzymes critical in nucleoside triphosphate (NTP) formation. AlgR2 positively regulates the production of alginate, GTP, ppGpp and inorganic polyphosphate (poly P). An algR2 mutant with low levels of these metabolites has them restored by introducing and overexpressing either the algR2 or the ndk gene into the algR2 mutant. Thus, Ndk is involved in the formation of these compounds and largely prevents the death of the algR2 mutant, which occurs early in the stationary phase. We demonstrate that the 12 kDa Ndk–pyruvate kinase (Pk) complex, previously shown to generate predominantly GTP instead of all the NTPs, has a low affinity for the deoxynucleoside diphosphates and cannot generate the dNTPs needed for DNA replication and cell division; this complex may thus be involved in regulating the levels of both NTPs and dNTPs that modulate cell division and survival in the stationary phase.

We report the purification and characterization of the enzyme nucleoside diphosphate kinase (Ndk) from Mycobacterium smegmatis. The N-terminus of the enzyme was blocked but an internal sequence showed approx. 70% homology with the same enzymes from Pseudomonas aeruginosa and Escherichia coli. immobilization of the mycobacterial nucleoside diphosphate kinase on a Sepharose 4 B matrix and passing the total cell extract through it revealed four proteins (P70, P65, P60, and P50, respectively) of M(r) 70 kDa, 65 kDa, 60 kDa and 50 kDa that were retained by the column. While the proteins of M(r) 70 kDa and 50 kDa modulated the activity of Ndk directing it towards GTP synthesis, the 60 kDa protein channelled the specificity of Ndk entirely towards CTP synthesis. The 65 kDa protein modulated the specificity of Ndk directing it entirely towards UTP synthesis. The specificity for such mycobacterial proteins towards NTP synthesis is retained when they are complexed with P. aeruginosa Ndk. We further demonstrate that the P70 protein is pyruvate kinase and that each of the four proteins forms a complex with Ndk and alters its substrate specificity. Given the ubiquitous nature of Ndk in the living cell and its role in maintaining correct ratios of intracellular nucleoside triphosphates, the implications of the occurrence of these complexes have been discussed in relation to the precursor pool for cell wall biosynthesis as well as RNA/DNA synthesis.

We have purified and characterized two class-II restriction endonucleases from the saprophyte Mycobacterium chelonei AMB 82. Mch AI was an isoschizomer of Not I recognizing and cleaving at 5'-GC/GGCCGC. MchA I did not require Triton X-100 for activity and was fully active at concentrations over 10–200 mM KCl and MgCl 2 . MchA II was an isoschizomer of Hae II recognizing and cleaving at 5'-GG/CC. MchA II was active in 10 mM Tris-HCl pH 8.0 containing MgCl 2 at 10–200 mM, was strongly inhibited by NaCl at concentrations higher than 10 mM and was inactive at KCl concentrations lower than 250 mM.

A restriction endonuclease, MgoI, an isoschizomer of Sau3AI, was purified from Mycobacterium gordonae TRC1318. As compared to Sau3AI, the yield of MgoI was seven-eightfold higher, the enzyme was four times more stable at 37 degrees C, and in addition, had optimal activity over a much broader range of salt concentrations.