Screening for cyclodextrin glycosyltransferase (CGTase)-producing alkaliphilic bacteria from samples collected from hyper saline soda lakes (Wadi Natrun Valley, Egypt), resulted in isolation of potent CGTase producing alkaliphilic bacterium, termed NPST-10. 16S rDNA sequence analysis identified the isolate as Amphibacillus sp. CGTase was purified to homogeneity up to 22.1 fold by starch adsorption and anion exchange chromatography with a yield of 44.7%. The purified enzyme was a monomeric protein with an estimated molecular weight of 92 kDa using SDS-PAGE. Catalytic activities of the enzyme were found to be 88.8 U mg⁻¹ protein, 20.0 U mg⁻¹ protein and 11.0 U mg⁻¹ protein for cyclization, coupling and hydrolytic activities, respectively. The enzyme was stable over a wide pH range from pH 5.0 to 11.0, with a maximal activity at pH 8.0. CGTase exhibited activity over a wide temperature range from 45 °C to 70 °C, with maximal activity at 50 °C and was stable at 30 °C to 55 °C for at least 1 h. Thermal stability of the purified enzyme could be significantly improved in the presence of CaCl₂. K_m and V_max values were estimated using soluble starch as a substrate to be 1.7 ± 0.15 mg/mL and 100 ± 2.0 μmol/min, respectively. CGTase was significantly inhibited in the presence of Co²⁺, Zn²⁺, Cu²⁺, Hg²⁺, Ba²⁺, Cd²⁺, and 2-mercaptoethanol. To the best of our knowledge, this is the first report of CGTase production by Amphibacillus sp. The achieved high conversion of insoluble raw corn starch into cyclodextrins (67.2%) with production of mainly β-CD (86.4%), makes Amphibacillus sp. NPST-10 desirable for the cyclodextrin production industry.

Compared to seeds and mature plants, sprouts are well characterized based on their nutritive values and biological properties. Moreover, laser light application is known to be a promising approach to improving plant growth, photosynthesis, and nutraceutical values. However, no studies have investigated the phytochemicals and biological activity of lemongrass (Cymbopogon proximus (Hochst. ex A.Rich.) Chiov.) sprouts or the further improvement of their quality by applying laser light treatment. We carried out a preliminary experiment for the optimization of laser treatment conditions, finding that a helium neon (He–Ne) laser at 632 nm and 5 mW for 5 min provided the most favorable conditions. We then investigated fresh weight, photosynthetic reactions, and primary and secondary metabolites, including sugars, amino acids, organic acids, essential oils, and phenolic compounds. Moreover, we studied the effect of laser light-induced changes in chemical compositions on the antioxidant, anti-diabetic, and anti-cholesterol activities of Cymbopogon proximus sprouts grown from laser-treated seeds. Laser light treatment increased the photosynthesis and respiration and hence the fresh weight of Cymbopogon proximus sprouts. Overall, sprouting increased most bioactive primary and secondary metabolites as compared to seeds. Increased photosynthesis by laser light improved carbon allocation and raised non-structural carbohydrates, which in turn led to improved synthesis of amino acids, organic acids, and essential oils, as well as phenolic and flavonoid compounds. As a result, laser light significantly improved the antioxidant capacity in terms of increasing the levels of ferric reducing antioxidant power (FRAP) (from 9.5 to 21 µmole trolox/g fresh weight (FW)), oxygen radical absorbance (ORAC) (from 400 to 1100 µmole trolox/100 g FW), and DPPH (from 5% to 25% of inhibation) and enhanced the hypocholesterolemic and antidiabetic activity through increasing the percentage of cholesterol micellar solubility (CMS) inhibition (from 42% to 62%) and glycemic index (from 33 to 17 µmole/g) over sprouts and seeds. In conclusion, the synergism of seed laser treatment and sprouting induced the health-promoting bioactive compounds in Cymbopogon proximus as compared to seeds, which can be applied at a large scale to improve the biochemical, physiological, and nutraceutical values of medicinal and crop sprouts.

Pelargonium graveolens, rose-scented geranium, is commonly used in the perfume industry. P. graveolens is enriched with essential oils, phenolics, flavonoids, which account for its tremendous biological activities. Laser light treatment and arbuscular mycorrhizal fungi (AMF) inoculation can further enhance the phytochemical content in a significant manner. In this study, we aimed to explore the synergistic impact of these two factors on P. graveolens. For this, we used four groups of surface-sterilized seeds: (1) control group1 (non-irradiated; non-colonized group); (2) control group2 (mycorrhizal colonized group); (3) helium-neon (He-Ne) laser-irradiated group; (4) mycorrhizal colonization coupled with He-Ne laser-irradiation group. Treated seeds were growing in artificial soil inculcated with Rhizophagus irregularis MUCL 41833, in a climate-controlled chamber. After 6 weeks, P. graveolens plants were checked for their phytochemical content and antibacterial potential. Laser light application improved the mycorrhizal colonization in P. graveolens plants which subsequently increased biomass accumulation, minerals uptake, and biological value of P. graveolens. The increase in the biological value was evident by the increase in the essential oils production. The concomitant application of laser light and mycorrhizal colonization also boosted the antimicrobial activity of P. graveolens. These results suggest that AMF co-treatment with laser light could be used as a promising approach to enhance the metabolic content and yield of P. graveolens for industrial and pharmaceutical use.

Herbal and traditional medicines can play a pivotal role in combating cancer and neglected tropical diseases. Ajuga bracteosa, family Lamiaceae, is an important medicinal plant. The genetic transformation of A. bracteosa with rol genes of Agrobacterium rhizogenes further enhances its metabolic content. This study aimed at undertaking the molecular, phytochemical, and in vitro biological analysis of A. bracteosa extracts. We transformed the A. bracteosa plant with rol genes and raised the regenerants from the hairy roots. Transgenic integration and expression of rolB were confirmed by conventional polymerase chain reaction (PCR) and qPCR analysis. The methanol: chloroform crude extracts of wild-type plants and transgenic regenerants were screened for in vitro antibacterial, antihemolytic, cytotoxic, anticancer, and leishmanial activity. Among all plants, transgenic line 3 (ABRL3) showed the highest expression of the rolB gene. Fourier transform infra-red (FTIR) analysis confirmed the enhanced number of functional groups of active compounds in all transgenic lines. Moreover, ABRL3 exhibited the highest antibacterial activity, minimum hemolytic activity (CC₅₀ = 7293.05 ± 7 μg/mL) and maximum antileishmanial activity (IC₅₀ of 56.16 ± 2 μg/mL). ABRL1 demonstrated the most prominent brine shrimp cytotoxicity (LD₅₀39.6 ± 4 μg/mL). ABRL3 was most effective against various human cancer cell lines with an IC₅₀ of 57.1 ± 2.2 μg/mL, 46.2 ± 1.1 μg/mL, 72.4 ± 1.3 μg/mL, 73.3 ± 2.1 μg/mL, 98.7 ± 1.6 μg/mL, and 97.1 ± 2.5 μg/mL against HepG2, LM3, A549, HT29, MCF-7, and MDA-MB-231, respectively. Overall, these transgenic extracts may offer a cheaper therapeutic source than the more expensive synthetic drugs.