Peptide Human Neutrophil Elastase Inhibitors from Natural Sources: An Overview

Elastases are a broad group of enzymes involved in the lysis of elastin, the main component of elastic fibres. They are produced and released in the human body, mainly by neutrophils and the pancreas. The imbalance between elastase activity and its endogenous inhibitors can cause different illnesses due to their excessive activity. The main aim of this review is to provide an overview of the latest advancements on the identification, structures and mechanisms of action of peptide human neutrophil elastase inhibitors isolated from natural sources, such as plants, animals, fungi, bacteria and sponges. The discovery of new elastase inhibitors could have a great impact on the pharmaceutical development of novel drugs through the optimization of the natural lead compounds. Bacteria produce mainly cyclic peptides, while animals provide for long and linear amino acid sequences. Despite their diverse natural sources, these elastase inhibitors show remarkable IC₅₀ values in a range from nM to μM values, thus representing an interesting starting point for the further development of potent bioactive compounds on human elastase enzymes.     

物理诱变法选育高产弹性蛋白酶的枯草芽胞杆菌

目的筛选弹性蛋白酶高产菌株,通过物理诱变,进一步提高弹性蛋白酶的产量。方法从土壤中收集样品,经反复初筛﹑复筛后,选择1株产弹性蛋白酶活性较高且稳定的细菌。通过对菌株的形态、生长特征及生理生化性质的研究,对菌株进行初步鉴定,然后进行紫外线诱变。结果该菌株属枯草芽孢杆菌(B1)。经诱变后得到1株诱变菌株,其HC值比原始菌株高1.33倍;酶活性达到120U/mL,比原始菌株提高了1.61倍。结论物理诱变可以有效提高弹性蛋白酶的酶活。     

菌源性弹性蛋白酶特性的研究

试验通过硫酸铵对发酵液的分段盐析得到粗酶，并对粗酶的基本性质进行了一定的研究。结果表明：酶反应的最适温度为50℃，最适pH为7．4；酶在37℃以下热稳定性良好；重金属对酶活性有不同程度的抑制作用；弹性蛋白对弹性蛋白酶具有较强的亲和吸附作用，弹性蛋白酶有优先水解弹性蛋白的能力。     

Kinetics of aspartame precursor synthesis catalysed by Pseudomonas aeruginosa elastase

Elastase isolated from Pseudomonas aeruginosa IFO 3455 was found to be an efficient protease to catalyse the synthesis of N-benzyloxycarbonyl-aspartyl-phenylalanine methyl ester, the precursor of the dipeptide sweetener, aspartame. The influence of methanol as a cosolvent in this synthetic reaction was investigated. It was found that the synthesis of the dipeptide precursor was most efficient in 25% (v/v) methanol, pH 7·0 at about 25°C for a reaction time of about 3 h. However, the activity of the enzyme was greatly reduced in 90% methanol. The values of K and k₂ for N-benzyloxycarbonyl-aspartic acid were 0·17 mol dm^?3 and 11·9 mol dm^?3 s^?1 respectively.     

枯草芽孢杆菌产弹性蛋白酶对肉嫩化的工艺研究

从前期研究枯草芽孢杆菌胞外分泌物中获得弹性蛋白酶,将之应用到猪肉背最长肌和牛主动脉,通过弹性蛋白酶溶液浸泡,采用嫩度计和电镜观察,发现此弹性蛋白酶作用于猪肉和主动脉,50℃,50min,添加量为4%,可以提高猪肉的嫩度。弹性蛋白酶的添加使牛主动脉的组织纹理打破,出现断裂和孔隙,可以使牛主动脉的韧度下降。     

Study of the refirming effect of a plant complex

Loss of skin elasticity is one of the main problems of ageing. This is a mechanical property influenced by elastin, a protein in the dermis which, together with collagen and glycosaminoglycans, makes up the connective tissue. This tissue is affected by a large number of events (such as cutaneous ageing, pregnancy, slimming processes and cellulitis) which eventually cause it to change. At the same time, the metabolism of the proteins of the connective tissue decreases and there is an ever greater presence of enzymes, principally elastases and collagenases, which are responsible for breaking down the elastin and the collagen. One way to prevent such a loss of elasticity is to use active ingredients that are able to inhibit elastase enzymes. A plant complex was prepared using the following plants: lady's thistle (Silybum marianum GAERTN), alchemilla or yarrow (Alchemilla vulgaris L.), horsetail (Equisetum arvense L.) as well as germinated seeds (Glycine soja Siebold and Zucc., Triticum vulgare Vilars, Medicago sativa L., Raphanus sativus L.). The complex was standardized to give the corresponding active principles, silybin, tannins, silicon and peptides, respectively, and in vitro enzymatic tests were carried out to establish its ability to inhibit elastase. The study of enzymatic inhibition was carried out using two enzymes: (1) porcine pancreatic elastase (PPE), and (2) human leukocyte elastase (HLE). The results showed that the plant complex presents non-competitive inhibition in the order of 41.0% against PPE and 50.0% against HLE. An in vivo test was made alongside the in vitro test using an SEM 474 Cutometer (Courage & Khazaka) to study the elasticity of the skin, and positive effects were obtained when applying a cosmetic formulation containing 5% of the plant complex. Image analysis of duplicates of the cutaneous surface, before and after treatment began with a product containing 5% of plant complex and showed that wrinkles were decreased by 36.7%.     

响应面优化弹性蛋白酶酶解罗非鱼下脚料的工艺研究

以罗非鱼加工鱼片的下脚料鱼头、鱼骨、鱼尾等为原料,选用弹性蛋白酶对其水解,通过响应面进行分析并优化酶解条件.以水解度与多肽含量为指标,分别对酶浓度、温度、pH值、料液比、酶解时间等因素对弹性蛋白酶酶解罗非鱼下脚料水解效果的影响进行分析.结果表明,弹性蛋白酶水解罗非鱼加工下脚料的最佳条件为：酶浓度0.553 mg/g,料液比1:5、自然pH、温度41.94 ℃、酶解3.31 h,在此条件下得到的水解度为18.09%,多肽含量为0.9 mg/mL.     

Protective and restorative effects of a Commiphora mukul gum resin and triheptanoin preparation on the CCL-110 skin fibroblast cell line

Coenzyme Q10 (CoQ10) is a major ingredient in skin care products because of its anti‐wrinkle effects, although it has some side effects especially at higher amounts. In this study, we compare the anti‐wrinkle related properties of CoQ10 and a proprietary Commiphora mukul gum resin (guggul) and triheptanoin preparation (GU‐TC7). GU‐TC7 is prepared with a supercritical CO₂‐co‐solvent extraction with ethanol, standardized to 2% guggulsterones and triheptanoin, a triglyceride composed of three 7‐carbon fatty acids. Treatment of CCL‐110 skin fibroblasts with GU‐TC7 demonstrates a mild proliferative effect compared to CoQ10 and increased type I collagen synthesis. Additionally, GU‐TC7 inhibited matrix metalloproteinase‐1 (MMP‐1) expression in a dose‐dependent manner at 20–100 μg mL^?1 and inhibited human elastase expression by more than 50% as compared to no elastase inhibition with CoQ10 treatment. These results suggest that GU‐TC7 possesses properties that are applicable to the treatment of wrinkles and may be considered for its further evaluation in skin care products.