Protamylasse, a residual compound of industrial starch production, provides a suitable medium for large-scale cyanophycin production

Protamylasse is a residual compound occurring during the industrial production of starch from potatoes. It contains a variety of nutrients and all necessary minerals and could be used as a carbon, nitrogen, and energy source for the growth of bacteria and also for cyanophycin (CGP) biosynthesis. Media containing protamylasse as the sole compound diluted only in water were therefore examined for their suitability in CGP production. Among various bacterial strains investigated in this study, a recombinant strain of Escherichia coli DH1 harboring plasmid pMa/c5-914::cphA6803, which carries the cyanophycin synthetase structural gene (cphA) from Synechocystis sp. strain PCC6803, was found to be most suitable. Various cultivation conditions for high CGP contents were first optimized in shake flask cultures. The optimized conditions were then successfully applied to 30- and 500-liter fermentation scales in stirred tank reactors. A maximum CGP content of 28% (wt/wt) CGP per cell dry matter was obtained in 6% (vol/vol) protamylasse medium at an initial pH of 7.0 within a cultivation period of only 24 h. The CGP contents obtained with this recombinant strain employing protamylasse medium were higher than those obtained with the same strain cultivated in mineral salts medium or in expensive commercial complex media such as Luria-Bertani or Terrific broth. It was shown that most amino acids present in the protamylasse medium were almost completely utilized by the cells during cultivation. Exceptions were alanine, tryptophan, tyrosine, and most interestingly, arginine. Furthermore, CGP was easily isolated from protamylasse-grown cells by applying the acid extraction method. The CGP exhibited a molecular mass of about 26 to 30 kDa and was composed of 50% (mol/mol) aspartate, 46% (mol/mol) arginine, and 4% (mol/mol) lysine. The use of cheap residual protamylasse could contribute in establishing an economically and also ecologically feasible process for the biotechnological production of CGP.     

Growth limitation and cell production bySaccharomycopsis fibuligera on a starch medium

Summary Batch cultures ofSaccharomycopsis fibuligera on a starch medium showed that the aerobic growth of the yeast was limited by total carbohydrate contents of the substrate, no inhibitory effect of metabolites being noticeable. Cell numbers increased earlier than dry weights of the biomass during adaptation to the medium. A significative increase in cell sizes accompanied active carbohydrate metabolism.     

Enhanced anthraquinones production from adsorbent-treated Morinda elliptica cell suspension cultures in production medium strategy

Continuous removal of anthraquinones (AQ) by Amberlite polymeric adsorbents (XAD-4, XAD-7 and XAD-16) through in situ adsorption in Morinda elliptica cell suspension cultures is studied for product recovery and improvement of the overall titre. Ethanol was the best eluting solvent for effective recovery of AQ from all adsorbents. Pre-treatment of XAD-4 with sodium acetate not only enhanced intracellular AQ, but also AQ release and subsequent recovery from the adsorbent. The addition of sodium acetate pretreated XAD-4 on day 18 for 6-day contact period, achieved comparable cell growth to control (41 g/L), but with 1.3-fold higher intracellular AQ (124 mg/g DW) and two-fold increase in extracellular AQ (14.3 mg/L). High amount of adsorbent and longer contact period for the cultures entering stationary growth phase, stimulated AQ release and recovery but at the expense of cell growth. With 5–8.3 g XAD-4 adsorbent per litre M. elliptica culture in production (P) medium, between 60 and 90% AQ was recovered from extracellular AQ after 24–26 days of culture period.     

Carotenoid production by Xanthophyllomyces dendrorhous: use of pineapple juice as a production medium

Aims:  To select a carotenoid-hyperproducing mutant and to formulate pineapple juice as a carotenoid production medium.
Methods and Results:  Yeast strain of Xanthophyllomyces dendrorhous mutant GM 807 (derived from gamma irradiation) and the mutant n78 (from neutron irradiation) were selected based on higher carotenoid production in diluted pineapple juice as a medium. The selected strain was evaluated under various concentrations of pineapple juice. The results obtained in the study demonstrate that the mutant GM 807 enhanced production by 65·8% when using pineapple juice as medium at 10% dilution in place of yeast malt medium.
Conclusion:  Pineapple juice could be used as a sole medium for carotenoid production.
Significance and Impact of the Study:  Our results provide useful information for the design of production media for carotenoids to be used in various applications.     

Experience with tarsal suspension as a factor in lower lid blepharoplasty

The bowed lower eyelid, with scleral show, is a common but untoward result following blepharoplasty with even minimal skin excision. A number of conditions, unrecognized preoperatively, can predispose a patient to scleral show. These include eyelid laxity with or without atrophic orbicularis muscle tone, lax canthal tendons, hypoplastic malar eminences, unrecognized Graves' ophthalmopathy, unilateral high myopia, or the secondary blepharoplasty. Suspension of the tarsus of the lower eyelid, concomitant with or following blepharoplasty, can straighten bowed lids and provide 2 to 3 mm of elevation, if desired. A classification of patients likely to develop scleral show is presented along with a revised technique of tarsal suspension.     

Preparation of cassava starch grafted with polystyrene by suspension polymerization

Cassava starch grafted with polystyrene (PS-g-starch) copolymer was synthesized via free-radical polymerization of styrene by using suspension polymerization technique. Potassium persulfate (PPS) was used as an initiator and water was used as a medium. The graft copolymer was characterized by Fourier transform infrared spectroscopy, differential scanning calorimetry, thermal gravimetric analysis, X-ray diffraction and scanning electron microscopy. The sub-micron spherical beads of PS were observed on the surface of starch granules. SEM micrographs showed porous patches of PS adhering on the starch granules after Soxhlet extraction. FTIR spectra also indicated the presence of PS-g-starch copolymer. XRD analysis exhibited insignificant changes in crystalline structure and degree of crystallinity. The effects of starch:styrene weight ratio, amount of PPS, reaction time and reaction temperature on the percentage of grafting – G (%), were investigated. G (%) increased with increasing starch content. Other variables showed their own individual optimal values. The optimum condition yielding 31.47% of G (%) was derived when the component ratio was 1:3 and reaction temperature and time were 50 °C and 2 h, respectively. Graft copolymerization did not change granular shape and crystallinity of starch. This study demonstrated the capability of polymerization of styrene monomer on the granular starch without emulsifier and the synthesis of graft copolymer without gelatinization of starch.     

Small grains for starch production

Wheat, sorghum, rice, barley, oat and rye grains are actual or potential raw materials for the industrial production of starch, but only the first three are so used. All six contain about 60% to 70% starch, and yield oil and protein as valuable byproducts of starch manufacture. Successful competition of these grains with the present major industrial sources of starch— corn, potatoes and cassava— depends on a number of factors, including comparative costs of the raw materials, efficiency of processing methods, and value of the byproducts.     

Post-thaw suspension of red cells cryopreserved with hydroxyethyl starch.

Post-thaw suspension of red cells cryopreserved with hydroxyethyl starch (HES) gives rise to an increase in the quantity of supernatant hemoglobin and a reduction in cell recovery. This occurs regardless of the suspension media used but may be delayed by some solutions. Extended suspension with most solutions results in loss of intracellular contents from a number of cells. The resulting ghosts account for 12–14% of the total cell population. In the presence of 14% HES, deposits on the surface of damaged cells indicate that cellular contents are adherent to the cell membrane and therefore do not contribute to the free hemoglobin in the external solution. The result is a misleading high cell recovery value and suggests that this test does not accurately describe true cell damage in the presence of HES. Since the saline stability of suspended cells changes very little from that prior to suspension (at thaw), its value is a more accurate assessment of cell damage.     

New medium composition for high betacyanin production by a cell suspension culture of table beet (Beta vulgaris L.)

The effect of a revised Linsmaier-Skoog (LS) medium on betacyanin production was investigated in suspension cultures of table beet (Beta vulgaris L.). The effects of a high iron concentration and low concentration of zinc on betacyanin production were not cumulative. The composition of the new revised medium for high betacyanin production was established by reducing the concentration of inorganic nitrogen (30 mM), modifying the ratio of ammonium to nitrate (1:14), reducing the concentration of zinc (0.0003 mM), and removing copper and cobalt. The revised LS medium enabled the maximum betacyanin yield of 550 mg/l to be obtained from a 14-day culture. This medium promoted the betacyanin production in three types of cell line differing in the betacyanin productivity. The betacyanin productivity (40 mg/l x day) was higher than that quoted in any other previous reports.     

Experience with studying suspension media in actinomycete lyophilization]

Viability, cultural features and antibiotic-production properties of the organisms producing tetracycline, chlortetracycline, erythromycin, neomycin, oxytetracycline and polymyxin were studied after their storage for 2 years in ampoules at a temperature of 4--10 degrees in lyophilized state with the use of sodium glutamate, polyvinylpyrrolidone, their combination and horse serum. The highest growth rate was observed in most of the cultures lyophilized in sodium glutamate. The growth of the cultures lyophilized in the solution of polyvinylpyrrolidone alone was mainly scanty or moderate. The antibiotic production level in some strains lyophilized in sodium glutamate or its combination with polyvinylpyrrolidone was after storage for 2 years somewhat higher than that in the control. The cultural features, i.e. the colour of the aerial and substrate mycelium and pigment secretion did not significantly differ in the lyophilized cultures and the cultures maintained on agarized media.     

Yeast hydrolysate as a low-cost additive to serum-free medium for the production of human thrombopoietin in suspension cultures of Chinese hamster ovary cells

To enhance the performance of a serum-free medium (SFM) for human thrombopoietin (hTPO) production in suspension cultures of recombinant Chinese hamster ovary (rCHO) cells, several low-cost hydrolysates such as yeast hydrolysate (YH), soy hydrolysate, wheat gluten hydrolysate and rice hydrolysate were tested as medium additives. Among various hydrolysates tested, the positive effect of YH on hTPO production was most significant. When 5 g l^–1 YH was added to SFM, the maximum hTPO concentration in batch culture was 40.41 g ml^–1, which is 11.5 times higher than that in SFM without YH supplementation. This enhanced hTPO production in YH-supplemented SFM was obtained by the combined effect of enhanced q_hTPO (the specific rate of hTPO production). The supplementation of YH in SFM increased q_hTPO by 294% and extended culture longevity by >2 days if the culture was terminated at a cell viability of 50%. Furthermore, cell viability throughout the culture using YH-supplemented SFM was higher than that using any other hydrolysate-supplemented SFM tested, thereby minimizing degradation of hTPO susceptible to proteolytic degradation. In addition, YH supplementation did not affect in vivo biological activity of hTPO. Taken together, the results obtained demonstrate the potential of YH as a medium additive for hTPO production in serum-free suspension cultures of rCHO cells.