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1.
A Panax notoginseng cell culture was successfully scaled up from shake flask to 1.0-L bubble column reactor and concentric-tube airlift reactor. High-density bioreactor batch cultivation was carried out using a modified MS medium. The maximum cell density in batch cultures reached 20.1, 21.0 and 24.1 g/L in the shake flask, bubble column and airlift reactors, respectively, and their corresponding biomass productivity was 950, 1140 and 1350 mg/(L x d) for each. The productivity of ginseng saponin was 70, 96 and 99 mg/(L x d) in the flask, bubble column and airlift reactors, respectively; and the polysaccharide productivity reached 104, 119 and 151 mg/(L x d) for each. Furthermore, a fed-batch cultivation strategy was developed on the basis of specific oxygen uptake rate (SOUR), i.e., sucrose feeding before a sharp decrease of SOUR, and the highest cell density of 29.7 g/L was successfully achieved in the airlift bioreactor on day 17 with a very high biomass productivity of 1520 mg/(L x d). The concentrations of ginseng saponin and polysaccharide reached about 2.1 and 3.0 g/L, respectively, and their productivity was 106 (saponin) and 158 mg/(L x d) (polysaccharide). This work successfully demonstrated the high-density bioreactor cultivation of P. notoginseng cells in pneumatically agitated bioreactors and the reproduction of the shake flask culture results in bioreactors. The cell density, biomass productivity, production titer and productivity of both ginseng saponin and polysaccharide obtained here were the highest that have been reported on a reactor scale for all the ginseng species. 相似文献
2.
Scale-up of a novel centrifugal impeller bioreactor (CIB) was demonstrated for production of valuable plant-specific secondary metabolites by high-density cell cultures. Initial kLa was identified to be a key factor affecting cell growth and production of ginseng saponin and polysaccharide by high-density cultivation of Panax notoginseng cells in a 3-L CIB. A high level of ginseng saponin and polysaccharide production was obtained at an initial kLa value of 30.2 h(-1). A maximum dry cell weight (DW) and production titer of ginseng saponin and polysaccharide reached 22.0 +/- 0.3, 1.5 +/- 0.1, and 2.7 +/- 0.2 g/L on day 15 with their corresponding productivity of 1140 +/- 42, 81 +/- 8, and 150 +/- 17 mg/(L.d), respectively. Based on initial kLa level, the CIB high-cell-density cultivation process was successfully scaled up from 3 L to 30 L. A maximum DW and production titer of ginseng saponin and polysaccharide in a 30-L CIB reached 25.5 +/- 0.5, 1.7 +/- 0.1, and 2.9 +/- 0.1 g/L (on day 15) at an initial kLa value of 28.7 h(-1), respectively, and their corresponding productivity was 1340 +/- 56, 91 +/- 9, and 164 +/- 15 mg/(L.d). Furthermore, by adopting a fed-batch cultivation strategy, a maximum DW and concentrations of total saponin and polysaccharide in the 30-L CIB were enhanced to 30.3 +/- 1.0, 2.1 +/- 0.1, and 3.5 +/- 0.2 g/L with their corresponding productivity of 1467 +/- 87, 102 +/- 13, and 179 +/- 18 mg/(L.d), respectively. The work suggests that the CIB may have great potential in large-scale high-density plant cell cultures for efficient production of useful secondary metabolites. 相似文献
3.
High cell density of Panax notoginseng in a 17 l airlift bioreactor was achieved in batch cultivation using a modified MS medium. The dry cell weight, ginseng saponin and polysaccharide reached 24, 1.7 and 2.8 g l–1, respectively, after 15 d. A strategy of sucrose feeding based on changes in the specific O2 uptake rate was applied to the cell cultures, which increased these respective yields to 30, 2.3 and 3.2 g l–1. 相似文献
4.
Production of ginseng and its bioactive components in plant cell culture: current technological and applied aspects 总被引:17,自引:0,他引:17
Ginseng (the root of Panax ginseng CA Mayer) is a valuable oriental herb, which has been used in traditional Chinese medicine for thousands of years, both as a disease-healing drug and a general tonic. The medicinal value of ginseng is now also widely recognized in the west and the world ginseng market is expanding. The current supply of ginseng depends mainly on field cultivation, which is a slow and laborious process. Plant cell and tissue culture methods have been explored as potentially more efficient alternatives for the mass production of ginseng and its bioactive components. Research into ginseng cell and tissue cultures started in the early 1960s and commercial applications have been underway since the late 1980s. The ginseng cell culture has continued to attract considerable research and development effort in recent years as scientists seek to understand and optimize the culture conditions. In this paper, we review recent studies on ginseng cell culture processes, focusing on the physiological and bioengineering factors affecting the productivity of ginseng biomass and useful metabolites (e.g. ginseng saponin and polysaccharide) and the progress and concerns in large-scale applications. 相似文献
5.
Effects of elicitation on the production of saponin in cell culture of Panax ginseng 总被引:1,自引:0,他引:1
This study was initiated to investigate the impacts of elicitor concentration and elicitor-adding time on the saponin synthesis and the cell growth of Panax ginseng cell suspensions. Both of the elicitors tested, yeast extract and methyl jasmonate, significantly improved saponin production. The highest additive level of the seven ginsenosides tested was 2.07% (dry weight basis), which was 28-fold higher than that in the control. The optimum time to add either elicitor was found to be on the day of inoculation. The addition of either elicitor did not show as significant an influence on cell growth as on saponin production. It was advisable to remove 2,4-dichlorophenoxyacetic acid (2,4-D) from the medium when methyl jasmonate was used as the elicitor as methyl jasmonate interacts antagonistically with 2,4-D. These results suggest that the addition of an elicitor to ginseng cell suspension cultures could stimulate saponin production. 相似文献
6.
We have established cell-suspension cultures of mountain ginseng (Panax ginseng G A. Mayer), and have attempted to increase the yield of saponin by manipulating our processing method and culturing factors
(e.g., media strengths; the presence of plant growth regulators or sucrose; ratios of NO+
3/ NH-
4). Maximum biomass yield was obtained in media containing 2,4-D. However, saponin productivity was much higher in a medium
comprising either IBA or NAA; 7.0 mg/L IBA was optimal for promoting both cell growth (10.0 g/L dry weight) and saponin production
(7.29 mg/g DW total ginsenoside). Although the addition of cytokinins (BA and kinetin) did not affect cell growth, the level
of saponin (particularly in the Rb group) was enhanced when the media were supplemented with either 0.5 mg/L BA or 0.5 mg/L
kinetin. Half- and full-strength MS media were equally suitable for inducing both biomass as well as saponin production. We
also investigated the effect of various concentrations of sucrose and nitrogen, and found that 30 g/L sucrose enhanced biomass
yield as well as saponin content However, further increases (i.e., up to 70 g/L) led to a decrease in saponin accumulation
and biomass production. Maximum growth and saponin productivity were reported from treatments with an initial nitrogen concentration
of 30 mM. In general, the amount of saponin increased when the test media had high NO+
3/ NH-
4 ratios; in fact, saponin production was greatest when nitrate was the sole nitrogen source. 相似文献
7.
Fungal elicitor induces singlet oxygen generation, ethylene release and saponin synthesis in cultured cells of Panax ginseng C. A. Meyer 总被引:1,自引:0,他引:1
Singlet oxygen is a high-energy molecular oxygen species. As one of the most active intermediates involved in chemical and biochemical reactions, singlet oxygen plays essential roles in plant responses to UV and strong light. Here, we report that Cle, an elicitor derived from fungal cell walls, induces the generation of singlet oxygen in cell cultures of ginseng, Panax ginseng. Cle treatment also triggers the activation of plasma membrane NADPH oxidase and 1-aminocyclopropane-1-carboxylic acid oxidase (ACO), subsequently leading to ethylene release and increased saponin synthesis, as shown by increased mRNA expression of squalene synthase (SQS) and squalene epoxidase (SQE), and accumulation of beta-amyrin synthase (beta-AS). Suppression of Cle-induced singlet oxygen generation or inhibition of ethylene production blocks saponin synthesis, whereas treatment of ginseng cells with ethylene or singlet oxygen induces the synthesis of saponin. Together, these results indicate that Cle-induced production of both singlet oxygen and ethylene is required for saponin synthesis, and that singlet oxygen may function upstream of ethylene during Cle-induced saponin synthesis. 相似文献
8.
T P Smolina T G Orlova O N Shcheglovitova N N Besednova 《Antibiotiki i khimioterapii͡a》1998,43(11):21-23
Induction of interferon (IF) and tumor necrosis factor (TNF) under the action of two polysaccharide preparations of ginseng i.e. panaxan-1 (from ginseng root) and panaxan-2 (from ginseng cell culture) was studied. Both the preparations induced production of TNF and IF in human leukocytes. By its properties and the typing results the induced IF proved to be gamma-IF. The preparation from the ginseng cell culture in the doses used had a higher IF inducing activity which could be explained by the difference in the polysaccharide composition of the preparations. 相似文献
9.
人参不同部位皂甙对衰老的人胚肺成纤维细胞影响的细胞化学研究 总被引:1,自引:0,他引:1
应用细胞化学的定性、定位,显微分光光度计定量分析的方法,测定人参不同部位皂甙对低代龄和高代龄人胚肺成纤维细胞内多糖类(PAS反应)、酸性非特异性酯酶(ANAE)、碱性磷酸酶(ALP)、酸性磷酸酶(ACP)、单胺氧化酶(MAO)含量的影响,实验结果表明:人参根、果、茎叶皂甙(SRG、SFG、SSLG)使低代龄细胞的多糖类、MAO含量下降或上升,对ANAE台量没有显著影响。但对高代龄细胞,SFG、SRG、SSLG则显著增加了多糖类、ANAE、ALP、ACP的相对含量,同时降低了MAO的含量。本文提示,人参不同部位皂甙均可以提高衰老细胞内多糖类,ALP、ACP、ANAE的相对含量,降低MAO的含量,而对年青细胞的影响则很不一致。 相似文献
10.
Jung JA Jhang E. John Staba Jung Yun Kim 《In vitro cellular & developmental biology. Plant》1974,9(4):253-259
Summary Roots, stems, or leaves of American (Panax quinquefolium) and Korean (Panax ginsing) ginseng were grown as callus or supension tissue cultures. Tissue cultures ofP. ginseng would occasionally form plantlets. The fundamental chemical composition, inorganic analysis, and saponin (panaquilin) content
of American and Korean ginseng plants and tissue cultures were determined. The crude saponin content is very similar to, but
approximately one-half (1.3%, fresh weight) of that present in ginseng roots. Two-dimensional thin layer chromatographic analysis
revealed minor differences in the panaquilins present in American and Korean ginseng tissue cultures. The sapogenin, panaxadiol,
was isolated from Korean ginseng callus. 相似文献
11.
Enhancement of growth and secondary metabolite biosynthesis: Effect of elicitors derived from plants and insects 总被引:1,自引:0,他引:1
Plant-derived natural products have been and will continue to be valuable sources. Elicitors have been employed to modify
cell metabolism in order to enhance the productivity of useful metabolites in plant cell/tissue cultures. In this study, several
elicitors were used to improve the productivity of useful metabolites and to reduce culture time for archiving high concentration
inP. ginseng hairy root cultures. The addition of chitosan, chitosan oligosaccharide and alginate oligosaccharide to the culture ofP. ginseng hairy roots caused growth to be inhibited with the increase in elicitor concentration. The usage of the chitosan elicitor
andd-glucosamine caused a slight decrease in hairy root growth, whereas total ginseng saponin accumulated slightly with the increase
in elicitor concentration. When gel beads were added to the culture medium at the initial period, hairy root growth was enhanced.
The maximum growth was 1.35 times higher than that of the control at 1% (w/v). Total ginseng saponin content decreased due
to the addition of alginate beads. This would result in consistent diffusion of lower levels of calcium ions during the culture
period that promotes biomass growth. 相似文献
12.
Juan Wang Wen-Yuan Gao Jian Zhang Bei-Mei Zuo Li-Ming Zhang Lu-Qi Huang 《In vitro cellular & developmental biology. Plant》2012,48(1):107-112
Based on the results of carbon source consumption in cell suspension culture of Panax quinquefolium L., 30 g L−1 sucrose was fed into a 5-L stirred tank bioreactor on day 16 of culture to enhance cell density and metabolite production.
Using a fed-batch cultivation strategy, polysaccharide production was enhanced to 1.608 g L−1, which was 1.96-fold greater than with batch cultivation. The maximum saponin yield (7.828 mg L−1) was obtained on day 24 and was about 36% higher than the yields obtained using batch cultivation. In a two-stage culture
process, a combined treatment with sucrose, lactoalbumin hydrolysate, and methyl jasmonate caused a significant increase in
total saponin yield (31.52 mg L−1) in cell cultures after 27 d. This value represents an increase of 4.03-fold compared with the total saponin yield in fed-batch
cultivation. The two-stage culture mode provided the best method for the in vitro production of secondary metabolites from P. quinquefolium. 相似文献
13.
模拟微重力环境因子对人参细胞生长和人参皂苷含量的影响 总被引:8,自引:1,他引:8
与在正常重力条件培养下的对照相比,经回转器水平回转处理的人参细胞鲜重和干重均增加,人参皂苷含量提高10%左右。在去Ca62+培养基上生长的人参愈伤组织细胞,经回转器水平回转3周后,人参皂苷含量约为正常重力条件下培养细胞的倍。另外,在试验范围内,如果培养基中直始钙离子浓度越高,则其培养的人参细胞中人参皂苷含量越低。 相似文献
14.
15.
Seung-Hyun Jo Suk-Yoon Kwon Doo-Sang Park Kyoung-Sil Yang Jae-Whune Kim Ki-Teak Lee Sang-Soo Kwak Haeng-Soon Lee 《Biotechnology and Bioprocess Engineering》2006,11(5):442-448
Human lactoferrin (hLf) is an iron-binding glycoprotein that has been considered to play many biological roles in the human,
including the stimulation of the immune system, antimicrobial and anti-inflammatory effects, and regulation of iron absorption.
We generated transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing a functional hLf protein using the signal peptide sequence from the endoplasmic reticulum and driven
by an oxidative stress-inducibleSWPA2 promoter which is highly expressed in plant cell cultures. The production of hLf increased proportionally to cell growth
and showed a maximal level (up to 3.6% of total soluble protein) at the stationary phase in suspension cultures. Full-length
hLf protein was identified by immunoblot analysis in transgenic cell cultures of Siberian ginseng. Recombinant hLf (rhLf)
was purified from suspension cells of Siberian ginseng by ammonium sulfate precipitation, cation-exchange and gel filtration
chromatography. N-terminal sequences of rhLf were identical to native hLf (nhLf). The overall monosaccharide composition of
rhLf showed the presence of plant specific xylose while sialic acid is absent. Antibacterial activity of purified rhLf was
higher than that of nhLf. Taken together, we anticipate that medicinal Siberian ginseng cultured cells, as demonstrated by
this study, will be a biotechnologically useful source for commercial production of functional hLf not requiring further purification. 相似文献
16.
西洋参与人参中人参皂甙含量的比较 总被引:2,自引:0,他引:2
采用TLC和HPLC方法分析比较西洋参(Panax quinquefolium L.)、人参(P.ginseng C.A.Mey.)及其加工品红参(red ginseng),以及不同规格的西洋参中人参皂甙的含量。结果表明,西洋参中人参皂甙总量及人参二醇型皂甙的含量明显高于人参及红参,且含有1种人参及红参中未发现的未知人参皂甙Rx,但不含人参及红参中含有的Rf;人参中人参二醇型皂甙的含量高于人参三醇 相似文献
17.
三七.人参和西洋参细胞悬浮培养的比较研究 总被引:19,自引:1,他引:18
用薄层层析对三七、人参和西洋参愈伤组织进行的初步鉴定表明,三种愈伤组织都含有皂甙和主要皂甙成分Rb_1、Rg_1,三七愈伤组织还含有一种抗癌皂甙Rh_1。对愈伤组织的生长,三七低于人参高于西洋参;对愈伤组织中总皂甙含量,三七均高于人参和西洋参。三种植物细胞悬浮培养结果类似于他们的愈伤组织培养,但生长又进一步提高。三七细胞悬浮培养中皂甙产生的时间进程几乎与生长平行,合适的收获期为培养30天。寡糖素不仅增强三七培养细胞的皂甙形成而且促进细胞生长,较合适的浓度为1.25 ppm。通过以上研究,使三七悬浮培养细胞的生长(干重增加178毫克)为最初培养愈伤组织的4倍以上,总皂甙产率高达20.6毫克,为最初培养愈伤组织的8.5倍。 相似文献
18.
19.
Rajesh Kumar Tewari Soo Yeon Lee Eun Joo Hahn Kee Yoeup Paek 《Plant biotechnology reports》2007,1(4):227-235
Nitric oxide (NO) is a diffusible, gaseous signaling molecule. In plants, NO influences growth and development, and it can
also affect plant responses to various stresses. Because NO induces root differentiation and interacts with reactive oxygen
species, we examined the temporal effect of NO elicitation on root growth, saponin accumulation and antioxidant defense responses
in the adventitious roots of mountain ginseng (Panax ginseng). The observations revealed that NO is involved in root growth and saponin production. Elicitation with sodium nitroprusside
(SNP) activated O2
−-generating NADPH oxidase (NOX) activity, which most probably subsequently enhanced growth of adventitious roots of mountain
ginseng. A severe inhibition of NOX activity and decline in dry weight of SNP elicited adventitious roots in the presence
of NOX inhibitor (diphenyl iodonium, DPI), which further supports involvement of NOX in root growth. Enhanced activities of
antioxidant enzymes by SNP appear to be responsible for low H2O2, less lipid peroxidation, and modulation of ascorbate and non-protein thiol statuses in the adventitious roots of mountain
ginseng. Dry mass, saponin content and NOX activity was related with NO content present in adventitious roots of mountain
ginseng. 相似文献
20.
与在正常重力条件培养下的对照相比,经回转器水平回转处理的人参细胞鲜重和干重均增加,人参皂苷含量提高10%左右。在去Ca2 培养基上生长的人参愈伤组织细胞,经回转器水平回转3周后,人参皂苷含量约为正常重力条件下培养细胞的2倍。另外,在试验范围内,如果培养基中起始钙离子浓度越高,则其培养的人参细胞中人参皂苷含量越低。 相似文献