不同营养方式对普通小球藻生长特性和细胞组成的影响

采用自养、异养和混合培养3种营养方式对普通小球藻进行了培养,分析了3种培养方式下小球藻的生长曲线、比生长速率和产率、培养基中pH和葡萄糖的变化,测定了藻细胞的光合色素、蛋白质和油脂含量及其产率。结果表明:混合营养条件下小球藻的比生长速率为自养的4.25～4.43倍、异养的0.78～1.00倍,产率为自养的5.79～6.27倍、异养的1.11～1.31倍;混合营养条件下,单位体积小球藻培养液中总叶绿素、蛋白质和油脂的产率分别为3.62 mg/(L·d)、53.41 mg/(L·d)和44.65 mg/(L·d),均高于自养和异养培养。研究认为,混合营养方式更加符合微藻生长的实际环境,是高密度、高含量活性物质培养微藻的理想方式,具有潜在的应用价值。     

培养条件对蛋白核小球藻生长及油脂合成的影响

以富油蛋白核小球藻为出发藻株,研究自养、异养和混养培养模式对小球藻生物量和油脂含量的影响,以及异养发酵培养基葡萄糖质量浓度、氮源种类及质量浓度对小球藻生长的影响。结果表明,与自养和混养培养模式相比,采用异养发酵方式培养蛋白核小球藻可获得最大的生物量和油脂含量。通过气相色谱法测得异养蛋白核小球藻油主要脂肪酸为棕榈酸(36.07%)、油酸(34.26%)、亚油酸(20.17%)和亚麻酸(6.12%)。经单因素试验优化得到最适蛋白核小球藻生长异养发酵培养基的葡萄糖质量浓度为60 g/L,最适的氮源为酵母粉,质量浓度为4 g/L,在此条件下经192 h发酵,蛋白核小球藻生物量可达12.43 g/L,油脂产量为5.45 g/L。研究结果表明,异养发酵培养获得的蛋白核小球藻油是一种潜在且可再生的新油源。     

异养小球藻主要营养成分及氨基酸组成分析

测定来自不同产地的4株蛋白核小球藻和2株椭圆小球藻异养培养后的主要营养成分和氨基酸组成,对其营养进行评价,并利用聚类和主成分分析探讨异养小球藻在营养成分和氨基酸含量上的差异。结果显示：异养小球藻主要营养成分均低于文献报导的自养小球藻,4株异养蛋白核小球藻和2株椭圆小球藻必需氨基酸占氨基酸总量的比例分别为34.94%～37.45%和37.70%～38.32%,必需氨基酸指数分别为37.50～49.52和131.89～135.77。影响异养蛋白核小球藻营养品质的限制性氨基酸为色氨酸、苏氨酸和异亮氨酸。聚类分析将4株异养蛋白核小球藻和2株异养椭圆小球藻各归为一类。主成分分析提取出两个主成分因子,主成分1和主成分2的贡献率分别为76.23%和17.80%,累积贡献率达94.03%。     

EMPLOYMENT OF ORGANIC ACIDS TO ENHANCE ASTAXANTHIN FORMATION IN HETEROTROPHIC CHLORELLA ZOFINGIENSIS

The green alga Chlorella zofingiensis produces significant amounts of the valuable ketocarotenoid astaxanthin under heterotrophic growth conditions. In this study, pyruvate, citrate and malic acid were investigated, for the first time, to promote the formation of secondary carotenoids including astaxanthin in C. zofingiensis in the dark. The addition of pyruvate, citrate and malic acid into the culture medium at a concentration above 10 mM stimulated biosynthesis of astaxanthin and other secondary carotenoids. The addition of 100 mM pyruvate enhanced the yield of astaxanthin from 8.36 to 10.72 mg/L, representing an increase of 28.2%. Citrate and malic acid also had stimulatory effects on the formation of astaxanthin. The results showed that organic acid supplementation was potentially useful for commercial production of astaxanthin by C. zofingiensis in dark heterotrophic culture.

PRACTICAL APPLICATIONS

This article revealed, for the first time, that the organic acids including pyruvate, citrate and malic acid could enhance the production of secondary carotenoids including the highly valuable product astaxanthin in the green alga Chlorella zofingiensis using fermentation technology. This research contributed significantly to our understanding of the synthesis of astaxanthin in green algae in dark heterotrophic cultures which would facilitate scale-up of the process to an industrial scale. In practice, organic acids could be used to effectively induce the formation of astaxanthin by C. zofingiensis during fermentation using organic carbon substrates as carbon sources, and at the same time to help adjust pH values favorably for astaxanthin production by C. zofingiensis in the fermenter.     

农杆菌介导小球藻快速转化方法的建立与条件优化

基于根瘤农杆菌（Agrobacterium tumefaciens）介导的方法实现异养蛋白核小球藻快速遗传转化,并优化转化条件。结果表明,小球藻异养生长条件的碳氮组合配比为葡萄糖20 g/L和酵母粉2 g/L时,生长速率为自养的3～5倍;根瘤农杆菌GV3101和LBA4404均可成功实现对异养小球藻的遗传转化,且转化效率无显著性差异（P＞0.05）。最优转化条件为根瘤农杆菌GV3101初始OD600 nm值= 0.8,小球藻（Chlorella pyrenoidosa）浓度为107个/mL,各取100 μL混合涂布在pH=5.4、乙酰丁香酮浓度为200 μmol/L的CM平板上,24 ℃避光培养40 h后转到筛选平板,仅2 d转化子便清晰可见,数目达88个/106微藻。转化过程整体耗时仅5 d,普遍比自养微藻转化时间缩短3倍以上,为小球藻代谢工程改造提供技术手段,同时为其他可异养培养微藻的短时高效遗传转化提供科学依据。     

MICROALGAE AS A SOURCE OF FATTY ACIDS

Microalgae are a very diverse group of organisms that consist of both prokaryotic and eukaryotic forms. Although most microalgae are phototrophic, some species are also capable of heterotrophic growth. Some species of microalgae can be induced to overproduce particular fatty acids through simple manipulations of the physical and chemical properties of the culture medium. As a result of the profound differences in cellular organization and growth modes and the ability to manipulate their fatty acid content, microalgae represent a significant source of unusual and valuable lipids and fatty acids.
The utilization of microalgal lipids and fatty acids as food components requires that these organisms be grown at large scale under controlled conditions. Several growth systems have been developed for large scale growth of phototrophic microalgae using either natural sunlight or artificial light, but their lack of control of culturing parameters or their high cost of operation have limited their utility for the production of food products. Instead, large scale cultivation of heterotrophic microalgae using classical fermentation systems provides consistent biomass produced under highly controlled conditions at low cost.
Microalgae contain many of the major lipid classes and fatty acids found in other organisms. However, they are also the principal producers in the biosphere of some polyunsaturated fatty acids, especially docosahexaenoic acid (DHA). The high DHA content found in some algae is currently being utilized to supplement infant formula to enable it to more closely resemble human breast milk. Thus, the commercialization of DHA from microalgae demonstrates the value of these organisms as a source of useful fatty acids.     

Algae in food: a general review

Abstract

Algae are common all over the Earth. Due to their rich chemical composition and content of bioactive substances they have been used in many fields of industry. Their gelling, thickening and stabilizing properties have led to the development of such products as agar, alginate and carrageenan. Moreover, algae are used in the food industry as food supplements and an addition to functional food. Algae are also added to meat products, such as pasty, steaks, frankfurters and sausages, as well as to fish, fish products, and oils, to improve their quality. Cereal-based products, such as pasta, flour and bread, are another group of products enriched with algae. Due to their properties algae may also be used for construction of fermented functional food. Fermented products containing algae are, most of all, dairy products, such as cheese, cream, milk deserts, yoghurt, cottage cheese, and processed cheese. Combination of fermented products offering a high content of lactic acid bacteria with algae possessing biologically active metabolites of natural origin allows not only to compose products with a high content of nutrients, but also to create a brand new segment of fermented food.     

开发营养丰富的海藻食品

海藻含有丰富的营养及特殊的生理活性成分,可广泛应用于海藻多糖、功能性食品、特效食品添加剂及保健制品等生产领域;其生产工艺有简单加工与深加工两种。海藻食品具有广阔的应用前景,据科学家预计,到下世纪初,藻类将为人类提供10％的蛋白质。 当今世界,生态及资源破坏日益严重,耕地减少及人口剧增引起的粮食短缺现象愈来愈紧迫;回归自然,享受绿色食品已成为全球的必然趋势,如何开发丰富的海洋食品资源,变得尤为重要。     

Nutritional Analysis of Puget Sound Bull Kelp (Nereocystis luetkeana)

Samples of Nereocystis luetkeana, an edible brown algae, were collected from Puget Sound, Wash. The freeze-dried samples were analyzed for proximate and elemental composition and evaluated for protein quality by amino acid analysis and using Tetrahymena pyriformis W. The freeze-dried frond, stipe, and bulb of the algae showed similar composition; however, the frond had a protein content twice that found in the bulb and stipe and five times less crude fiber than the stipe. The proximate analysis of the frond was 15.3% protein, 1.3% crude fiber, 42.7% ash, 1.9% crude fat, and 38.8% carbohydrate (by difference). Based on neutron activation analysis, Nereocysti: contained appreciable quantities of sodium, potassium, calcium, and iron. The algae protein appeared to be of high quality based on growth of Tetrahymena pyriformis W. on a pepsin digest of the algae and on the amino acid profile and in vitro digestibility of the intact algae.     

氮浓度对异养小球藻生长及蛋白质含量的影响

为了提高异养小球藻蛋白质含量低的问题,研究了异养小球藻生长及蛋白质含量和氮浓度的关系,探究了分阶段调控氮浓度对异养小球藻生长及蛋白质含量的影响。结果表明,氮浓度在3～15mmol/L范围内,小球藻生物量及蛋白质含量随氮浓度增加而增加,生物量从0.91g/L提高到了3.02g/L,蛋白质含量从26.1%提高到了37.4%。分阶段培养小球藻,首先在低氮条件下培养至指数期前期,然后转移至高氮浓度下培养,生物量达3.04g/L,且蛋白质含量提高至53.8%,与自养培养条件下蛋白质含量相当。     

Effects of dietary supplements of algae,containing polyunsaturated fatty acids,on milk yield and the composition of milk products in dairy ewes

Thirty-two lactating Karagouniko ewes were allocated at random to four groups for 6 weeks, to examine the effect of four diets: C (control treatment, ration without algae); LA (ration with low level of algae); MA (ration with medium level of algae) and HA (ration with high level of algae); containing 0, 23.5, 47 and 94 g algae, respectively, on the enrichment of milk and dairy products. Addition of algae reduced (P < 0.001) DM intake for treatments MA and HA. Milk yield did not differ between treatments but milk composition was significantly affected by dietary inclusion of algae. Milk fat content was significantly increased (P < 0.001) for treatment HA whereas milk protein content was significantly increased (P < 0.001) for all treatments containing algae. Milk from treatments LA, MA and HA was significantly enriched in the following PUFA: C20:5 (n-3) (0.4-2.1%), C22:5 (n-6) (0.8-4.1%), C22:6 (n-3) (4.3-12.4%) (P < 0.001) and C22:5 (n-3) (2.1-3.1%) (P < 0.05), which were not detected in control milk. Feta cheese and yogurts produced from the enriched milk had identical composition with the milk, and would be characterized as healthy foods. The ratio of n-6 to n-3 fatty acids was 2.5-4.5.     

Short communication: The effect of substituting fish oil in dairy cow diets with docosahexaenoic acid-micro algae on milk composition and fatty acids profile

The effects of substituting fish oil (FO) with docosahexaenoic acid (DHA)-micro algae on milk chemical and fatty acid composition were examined in this study. Twenty-four Holstein cows in mid lactation grazing on an alfalfa-grass based pasture were divided into 4 treatment groups (6 cows/treatment) and supplemented with 7 kg/d grain mix plus 350 g of soybean oil and one of the following: 1) 150 g of FO, 2) 100 g of FO plus 50 g of algae, 3) 50 g of FO plus 100 g of algae, or 4) 150 g of algae. Cows were fed treatment diets for 3 wk, and milk samples were collected from each cow during the last 3 d of the study. Milk production (17.96, 17.56, 17.55, and 19.26 kg/d for treatment diets 1 to 4, respectively), milk fat percentages (3.17, 3.49, 3.74, and 3.43%), and milk protein percentages (3.35, 3.50, 3.71, and 3.42%) were similar between treatment diets. Concentrations (g/100 g of fatty acids) of milk cis-9 trans-11 (c9t11) conjugated linoleic acid (CLA; 3.41, 3.69, 4.47, and 4.21 for treatment diets 1 to 4, respectively) and vaccenic acid (11.80, 12.83, 13.87, and 13.53) were similar between treatment diets. Results of this study suggest that DHA-micro algae can partially or fully substitute FO in a cow's diet without any adverse effects on milk production, milk composition, or milk c9t11 CLA content. The DHA-micro algae may be used as a viable alternative for FO in cow's diet to modify rumen biohydrogenation to increase milk c9t11 CLA content.     

A steady-state biofilm model for simultaneous reduction of nitrate and perchlorate, part 1: model development and numerical solution

A multispecies biofilm model is developed for simultaneous reduction of nitrate and perchlorate in the H(2)-based membrane biofilm reactor. The one-dimension model includes dual-substrate Monod kinetics for a steady-state biofilm with five solid and five dissolved components. The solid components are autotrophic denitrifying bacteria, autotrophic perchlorate-reducing bacteria, heterotrophic bacteria, inert biomass, and extracellular polymeric substances (EPS). The dissolved components are nitrate, perchlorate, hydrogen (H(2)), substrate-utilization-associated products, and biomass-associated products (BAP). The model explicitly considers four mechanisms involved in how three important operating conditions (H(2) pressure, nitrate loading, and perchlorate loading) affect nitrate and perchlorate removals: (1) competition for H(2), (2) promotion of PRB growth due to having two electron acceptors (nitrate and perchlorate), (3) competition between nitrate and perchlorate reduction for the same resources in the PRB: electrons and possibly reductase enzymes, and (4) competition for space in the biofilm. Two other special features are having H(2) delivered from the membrane substratum and solving directly for steady state using a novel three-step approach: finite-difference for approximating partial differential and/or integral equations, Newton-Raphson for solving nonlinear equations, and an iterative scheme to obtain the steady-state biofilm thickness. An example result illustrates the model's features.     

Effect of glycine on 5-aminolevulinic acid biosynthesis in heterotrophic culture of Chlorella regularis YA-603

Chlorella regularis strain YA-603 was previously found to exhibit a comparatively high growth rate and improved 5-aminolevulinic acid (ALA) productivity when cultured heterotrophically. Although the universal tetrapyrrole precursor ALA is synthesized from glutamate in algae and higher plants, in this study the addition of glutamate to the medium did not enhance ALA production in the heterotrophic culture of C. regularis YA-603. On the other hand, the addition of glycine, which is a precursor of ALA biosynthesis via the Shemin pathway in animal cells and some bacteria, enhanced both the specific growth rate and ALA production. Moreover, when a second glycine addition was made during the culture, the ALA concentration increased by about 1.5 times compared to that obtained with one glycine addition. From these results, it is suggested that the Shemin pathway contributes to ALA production, and that addition of glycine in the heterotrophic culture of C. regularis YA-603 can significantly increase the amount of ALA produced.     

Identification of selected seaweed polysaccharides (phycocolloids) by vibrational spectroscopy (FTIR-ATR and FT-Raman)

The wide industrial application of phycocolloids (e.g. alginates, agar and carrageenans) is based on their particular properties to form gels in aqueous solution. These seaweed polysaccharides present a chemical structure related with the taxonomic position of the algae: carrageenans are produced by carrageenophytes (red algae belonging mainly to the genera Kappaphycus, Eucheuma, Chondrus, Gigartina and Chondracanthus). Recently, new spectroscopic techniques have provided more accurate identification of the natural composition of the polysaccharides produced by these seaweeds. With the combination of two spectroscopic techniques (FTIR-ATR and FT-Raman) it is possible to identify the principal seaweed colloids in ground seaweed samples as in extracted material. Since the seaweed samples receive the minimum of handling and treatment (e.g. they are simply dried and ground), the composition determined represents, as accurately as possible, the native composition of the phycocolloids.     

Lipid,fatty acid,protein, amino acid and ash contents in four Brazilian red algae species

Four species of marine benthic algae (Laurencia filiformis, L. intricata, Gracilaria domingensis and G. birdiae) that belong to the phylum Rhodophyta were collected in Espírito Santo State, Brazil and investigated concerning their biochemical composition (fatty acid, total lipid, soluble proteins, amino acid and ash). The total content of lipid (% dry weight) ranged from 1.1% to 6.2%; fatty acid from 0.7% to 1.0%; soluble protein from 4.6% to 18.3%, amino acid from 6.7% to 11.3% and ash from 22.5% to 38.4%. Judging from their composition, the four species of algae appear to be potential sources of dietary proteins, amino acids, lipids and essential fatty acids for humans and animals.     

Hydrogen sulfide oxidation by a microbial consortium in a recirculation reactor system: sulfur formation under oxygen limitation and removal of phenols

Wastewater from petroleum refining may contain a number of undesirable contaminants including sulfides, phenolic compounds, and ammonia. The concentrations of these compounds must be reduced to acceptable levels before discharge. Sulfur formation and the effect of selected phenolic compounds on the sulfide oxidation were studied in autotrophic aerobic cultures. A recirculation reactor system was implemented to improve the elemental sulfur recovery. The relation between oxygen and sulfide was determined calculating the O2/S2- loading rates (Q(O2)/Q(S)2- = Rmt), which adequately defined the operation conditions to control the sulfide oxidation. Sulfur-producing steady states were achieved at Rmt ranging from 0.5 to 1.5. The maximum sulfur formation occurred at Rmt of 0.5 where 85% of the total sulfur added to the reactor as sulfide was transformed to elemental sulfur and 90% of it was recovered from the bottom of the reactor. Sulfide was completely oxidized to sulfate (Rmt of 2) in a stirred tank reactor, even when a mixture of phenolic compounds was present in the medium. Microcosm experiments showed that carbon dioxide production increased in the presence of the phenols, suggesting that these compounds were oxidized and that they may have been used as carbon and energy source by heterotrophic microorganisms present in the consortium.     

Microbial ecology of nitrifying bacteria in wastewater treatment process examined by fluorescence in situ hybridization

The microbial ecology of nitrifying bacteria in various types of wastewater treatment processes and the dynamic response of the microbial ecology in biofilms were investigated using fluorescence in situ hybridization (FISH) with 16S rRNA-targeted oligonucleotide probes. Nitrifying bacteria were found to exhibit various organizational forms under different conditions of substrate composition and concentration. Ammonia-oxidizing bacteria were dominant in ammonia-rich inorganic wastewater, while heterotrophic bacteria and ammonia-oxidizing bacteria were localized at different positions in the biofilm in organic wastewater. The dynamics of the microbial ecology in the biofilm with regard to the spatial distribution of ammonia-oxidizing bacteria and heterotrophic bacteria caused by a gradual change in substrate composition was successfully monitored by FISH analysis.     

Heterotrophic plate count monitoring of treated drinking water in the UK: a useful operational tool

The count of general or heterotrophic bacterial populations in treated drinking water in the UK has been undertaken since the 1880s. Counts of heterotrophic bacteria at 22 and 37 degrees C are used widely as part of an overall assessment of treated drinking water quality. There were no legislated standards for water quality in the UK until adoption of the first EU Directive in 1989. The UK has, however, never stipulated numerical standards or guidelines for heterotrophic bacteria, although their enumeration has long been part of the assessment of 'wholesome' water, on which advice regarding microbiological quality was given in a series of documents known as 'Report 71'. The current regulations stipulate only that there should be 'no abnormal change' in numbers normally associated with a given supply. This paper reviews the historical context regarding the enumeration, and interpretation of results, of heterotrophic bacteria from treated drinking water, and information regarding current practices by UK water suppliers. The appropriateness of using heterotrophic bacteria counts as an operational tool or as a health parameter is briefly discussed in the light of the UK experience.     

植物化感作用在控制水华藻类中的应用

伴随着水环境污染,水体中藻类疯长形成的"水华"、"赤潮"现象日益严重。各种化学、物理、生物方法被用于抑制藻类的生长,但这些方法都存在不易控制、成本高、易破坏生态等问题。利用植物化感作用抑制藻类生长具有生态安全和灵敏高效等优点,对湖泊富营养化的生态控制具有非常重要的意义。作者介绍了植物化感抑藻的研究进展,归纳了化感作用的种类和化感物质的化学成分,并讨论了植物化感抑藻的作用机制、抑藻机理,最后对植物化感作用在抑藻方面的研究前景进行了展望。