Search

Flowable resins used for dental restoration are subject to biofilm formation. Zinc has antibacterial properties. Thus, we prepared a zinc-doped phosphate-based glass (Zn-PBG) to dope a flowable resin and evaluated the antibacterial activity of the composite against Streptococcus mutans (S. mutans) to extrapolate the preventative effect toward secondary caries. The composites were prepared having 0 (control), 1.9, 3.8, and 5.4 wt.% Zn-PBG. The flexural strength, elastic modulus, microhardness, depth of cure, ion release, inhibition zone size, and number of colony-forming units were evaluated and analyzed using ANOVA. The flexural strength of the control was significantly higher than those of Zn-PBG samples (p &lt, 0.05). However, all samples meet the International Standard, ISO 4049. The microhardness was not significantly different for the control group and 1.9 and 3.8 wt.% groups, but the 5.4 wt.% Zn-PBG group had a significantly lower microhardness (p &lt, 0.05). Further, the composite resins increasingly released P, Ca, Na, and Zn ions with an increase in Zn-PBG content (p &lt, 0.05). The colony-forming unit count revealed a significant reduction in S. mutans viability (p &lt, 0.05) with increase in Zn-PBG content. Therefore, the addition of Zn-PBG to flowable composite resins enhances antibacterial activity and could aid the prevention of secondary caries.

A two-stage enzymatic process was developed for agarose hydrolysis without acid pretreatment. In the first stage, agarose was hydrolyzed to produce neoagarobiose using an optimized dosage of AgaG1 and DagB at pH 7.0 and 40 °C. Agarose gelation was avoided by momentarily elevating the reaction temperature for the first 10 min, instead of employing a pretreatment step with acid prior to the enzyme reaction. In the second stage, neoagarobiose was further hydrolyzed to produce galactose using neoagarobiose hydrolase (NABH) at pH 7.0 and 35 °C. The overall yield of galactose from agarose was 88% of the theoretical maximum. The crude galactose solution produced from agarose hydrolysis was used directly for ethanol production by yeast to demonstrate its potential. In overall, 20 g/l of agarose could be converted to 3.71 g/l of ethanol.

Alteromonas sp. GNUM-1 is known to degrade agar, the main cell wall component of red macroalgae, for their growth. A putative agarase gene (agaG1) was identified from the mini-library of GNUM-1, when extracellular agarase activity was detected in a bacterial transformant. The nucleotide sequence revealed that AgaG1 had significant homology to GH16 agarases. agaG1 encodes a primary translation product (34.7 kDa) of 301 amino acids, including a 19-amino-acid signal peptide. For intracellular expression, a gene fragment encoding only the mature form (282 amino acids) was cloned into pGEX-5X-1 in Escherichia coli, where AgaG1 was expressed as a fusion protein with GST attached to its N-terminal (GST-AgaG1). GST-AgaG1 purified on a glutathione sepharose column had an apparent molecular weight of 59 kDa on SDS-PAGE, and this weight matched with the estimated molecular weight (58.7 kDa). The agarase activity of the purified protein was confirmed by the zymogram assay. GST-AgaG1 could hydrolyze the artificial chromogenic substrate, p-nitrophenyl-β-d-galactopyranoside but not p-nitrophenyl-α-d-galactopyranoside. The optimum pH and temperature for GST-AgaG1 activity were identified as 7.0 and 40 °C, respectively. GST-AgaG1 was stable up to 40 °C (100 %), and it retained more than 70 % of its initial activity at 45 °C after heat treatment for 30 min. The K m and V max for agarose were 3.74 mg/ml and 23.8 U/mg, respectively. GST-AgaG1 did not require metal ions for its activity. Thin layer chromatography analysis, mass spectrometry, and 13C-nuclear magnetic resonance spectrometry of the GST-AgaG1 hydrolysis products revealed that GST-AgaG1 is an endo-type β-agarase that hydrolyzes agarose and neoagarotetraose into neoagarobiose.

Heterologous expression of a putative K+/H+ antiporter of Streptomyces coelicolor A3(2) (designated as sha4) in E. coli and Streptomyces hygroscopicus JCM4427 showed enhanced tolerance to K+ stress, acidic-pH shock, and/or geldanamycin production under K+ stress. In a series of K+ extrusion experiments with sha4-carrying E. coli deficient in the K+/H+ antiporter, a restoration of impaired K+ extrusion activity was observed. Based on this, it was concluded that sha4 was a true K+/H+ antiporter. In different sets of experiments, the sha4-carrying E. coli showed significantly improved tolerances to K+ stresses and acidic-pH shock, whereas sha4-carrying S. hygroscopicus showed an improvement in K+ stress tolerance only. The sha4-carrying S. hygroscopicus showed much higher geldanamycin productivity than the control under K+ stress condition. In another set of experiments with a production medium, the secretion of geldanamycin was also significantly enhanced by the expression of sha4.