Identification of Bioactive Peptides from a Laminaria digitata Protein Hydrolysate Using In Silico and In Vitro Methods to Identify Angiotensin-1-Converting Enzyme (ACE-1) Inhibitory Peptides.

Bioactive peptides range in size from 2-30 amino acids and may be derived from any protein-containing biomass using hydrolysis, fermentation or high-pressure processing. Pro-peptides or cryptides result in shorter peptide sequences following digestion and may have enhanced bioactivity. Previously, we identified a protein hydrolysate generated from Laminaria digitata that inhibited ACE-1 in vitro and had an ACE-1 IC50 value of 590 µg/mL compared to an ACE-1 IC50 value of 500 µg/mL (~2.3 µM) observed for the anti-hypertensive drug Captopril©. A number of peptide sequences (130 in total) were identified using mass spectrometry from a 3 kDa permeate of this hydrolysate. Predicted bioactivities for these peptides were determined using an in silico strategy previously published by this group utilizing available databases including Expasy peptide cutter, BIOPEP and Peptide Ranker. Peptide sequences YIGNNPAKGGLF and IGNNPAKGGLF had Peptide Ranker scores of 0.81 and 0.80, respectively, and were chemically synthesized. Synthesized peptides were evaluated for ACE-1 inhibitory activity in vitro and were found to inhibit ACE-1 by 80 ± 8% and 91 ± 16%, respectively. The observed ACE-1 IC50 values for IGNNPAKGGLF and YIGNNPAKGGLF were determined as 174.4 µg/mL and 133.1 µg/mL. Both peptides produced sequences following simulated digestion with the potential to inhibit Dipeptidyl peptidase IV (DPP-IV).

Angiotensin-I-Converting Enzyme Inhibitory Activity of Protein Hydrolysates Generated from the Macroalga Laminaria digitata (Hudson) JV Lamouroux 1813.

Seaweeds have a long history of use as both food and medicine, especially in Asian cultures. Moreover, there is growing interest in the use of seaweed ingredients and bioactive compounds in pharmaceutical and nutraceutical products. One ailment that seaweed bioactive compounds may impact is hypertension caused by the enzyme Angiotensin Converting Enzyme 1 (ACE-1; EC 3.4.15.1), found within the Renin-Angiotensin Aldosterone System (RAAS), which causes vasoconstriction of blood vessels, including veins and arteries. The aim of this paper is to generate bioactive peptide containing protein hydrolysates from the brown seaweed Laminaria digitata (Hudson) JV Lamouroux 1813. Proteins were extracted from this seaweed by disrupting the seaweed cell wall using a combination of carbohydrases and proteolytic enzymes. Bioactive peptide containing permeates were generated from L. digitata protein hydrolysates, and both hydrolysates and permeates were screened for their ability to inhibit the enzyme ACE-1. The protein content of the permeate fractions was found to be 23.87% compared to the untreated seaweed, which contained 15.08% protein using LECO analysis. Hydrolysis and filtration resulted in a "white" protein powder, and the protein content of this powder increased by 9% compared to the whole seaweed. The total amino acid (TAA) content of the L. digitata protein permeate was 53.65 g/100 g of the sample, and contains over 32% essential amino acids (EAA). Furthermore, the L. digitata permeate was found to inhibit the ACE-1 enzyme by 75% when compared to the commercial drug Captopril© when assayed at a concentration of 1 mg/mL. The inhibition of ACE-1 (the IC50 value) of 590 µg/mL for the L. digitata permeate compares well with Captopril©, which had 100% inhibition of ACE-1, with an IC50 value of 500 µg/mL. This study indicates that there is potential to develop protein powders with ACE-1 inhibitory bioactivities from the brown seaweed L. digitata using enzymatic hydrolysis as a cell disruption and protein extraction/hydrolysate generation procedure.

Carbonaceous and nitrogenous disinfection by-product formation from algal organic matter.

Seasonal algal blooms in drinking water sources release intracellular and extracellular algal organic matter (AOM) in significant concentrations into the water. This organic matter provides precursors for disinfection by-products (DBPs) formed when the water is subsequently chlorinated at the final disinfection stage of the potable water treatment process. This paper presents results of AOM characterisation from five algal species (three cyanobacteria, one diatom and one green) alongside the measurement of the DBP formation potential from the AOM of six algal species (an additional diatom). The character was explored in terms of hydrophilicity, charge and protein and carbohydrate content. 18 DBPs were measured following chlorination of the AOM samples: the four trihalomethanes (THMs), nine haloacetic acids (HAAs), four haloacetonitriles (HANs) and one halonitromethane (HNM). The AOM was found to be mainly hydrophilic (52 and 81%) in nature. Yields of up to 92.4 µg mg-1 C carbonaceous DBPs were measured, with few consistent trends between DBP formation propensity and either the specific ultraviolet absorbance (SUVA) or the chemical characteristics. The AOM from diatomaceous algae formed significant amounts of nitrogenous DBPs (up to 1.7 µg mg-1 C). The weak trends in DBPFP may be attributable to the hydrophilic nature of AOM, which also makes it more challenging to remove by conventional water treatment processes.

The influence of ultrasound frequency and power, on the algal species Microcystis aeruginosa, Aphanizomenon flos-aquae, Scenedesmus subspicatus and Melosira sp.

We report on the effectiveness of sonication on controlling the growth of four problematic algal species which are morphologically different and from three algal divisions. Two cyanobacterial species Microcystis aeruginosa (unicellular) and Aphanizomenon flos-aquae (filamentous), one green alga Scenedesmus subspicatus (colonial) and lastly a diatom species Melosira sp. (filamentous) were subjected to ultrasound of selected low to high frequencies ranging from 20 to 1144 kHz. Microcystis aeruginosa and Scenedesmus subspicatus highest cell removal rates were 16 +/- 2% and 20 +/- 3% when treated with the same ultrasound frequency of 862 kHz but differing energy levels of 133 and 67 kWh m(-3), respectively. Aphanizomenon flos-aquae best removal rate was 99 +/- 1% after 862 kHz and 133 kWh m(-3) of energy, with Melosira sp. achieving its highest cell removal at 83% subsequent to ultrasound of 20 kHz and 19 kWh m(-3). Microcystis aeruginosa and Scenedesmus subspicatus are considered non-susceptible species to ultrasound treatment from a water treatment perspective due to their low cell removal rates; however, photosynthetic activity reduction of 65% for Microcystis aeruginosa does indicate the possible utilization of ultrasound to control bloom growth, rather than bloom elimination. Conversely, Aphanizomenon flos-aquae and Melosira sp. are deemed species highly susceptible to ultrasound. Morphological differences in shape (filamentous/non-filamentous) and cell wall structure (silica/peptidoglycan), and presence of gas vacuoles are probable reasons for these differing levels of susceptibility to ultrasound.

The effects of temperature, pH and sulphide on the community structure of hyperthermophilic streamers in hot springs of northern Thailand.

Hyperthermophilic community diversity was assessed in hot-spring streamers along gradients of temperature, pH and sulphide in northern Thailand. A hierarchical sampling design was employed to obtain biomass for culture-independent estimates of 16S rRNA gene-defined prokaryotic diversity. All springs supported several archaeal and bacterial phylotypes, including novel phylotypes that expand the known phylogenetic diversity of terrestrial hyperthermophiles. Diversity appeared significantly greater than that observed for several other geographic locations. Phylotypes belonging to the Aquificales were ubiquitous, further supporting the hypothesis that these chemolithoautotrophs are key members of all hyperthermophilic communities. The chemoorganotrophic genus Thermus was also represented by phylotypes in all springs. Other bacterial taxa represented by environmental sequences included Bacillus, Thermotoga and various unidentified Alphaproteobacteria and Betaproteobacteria. Archaeal phylotypes included the Crenarchaea Desulfurococcus, Pyrobaculum, plus several unidentified hyperthermophilic lineages. A Methanothermococcus-like Euryarchaeon was also identified, with this genus not previously known from streamer communities. A multivariate approach to the analysis of biotic and abiotic data revealed that diversity patterns were best explained by a combination of temperature and sulphide rather than by any other abiotic variable either individually or in combination.