CS270332B1 - Method of yeast products of methanol production - Google Patents

Abstract

The workbook deals with the production of yeast biomass from methanol, which is rich in protein, essential amino acids and vitamins and that serves as a valuable protein and vitamin food for farm animals and as a source for the preparation of Cl specific enzymes Like alcohol oxidase, formaldehyde- and formate dehydrogenase, dihydroxyacetone synthase, further catalase, NADH, GSII and ATP from methanol. "Yeast Production Method." products yeast biomass from methanol, increase concentration of cells in the medium and shorten the time cultivation. In the production of yeast products Methylotrophic from methanol are used especially yeast strains of Candida boidinii 2 and Hansenula ep.3 with culture medium contains, in addition to methanol, 0.5 to 1% by weight of molasses distillers in an amount of 0.2 to 0.3% by weight at a temperature of 30 to 40 ° C and pH 3.0 up to 3.5.

Description

EN 270332 B1 1

The invention relates to a process for the production of yeast products from methanol.

Molasses distillers are commonly used to produce feed yeast. They are a precursor, not a carbon source, a large amount of potassium, assimilable nitrogen, and sometimes enough magnesium. What is important is the content of the growth filaments that come into the stillage from beet molasses, but also from yeast, during the fermentation of the distillery, and the presence of a wide range of trace elements. The disadvantage of molasses distillers as a raw material for the production of feed yeast proteins is their uneven composition, low carbon content and the need to add large amounts of acids to their use in adjusting the pH of the fermentation broth during cultivation. Therefore, molasses distillers are preferably used as an addition to the medium in the production of yeast proteins from sulphite extracts, from ethanol-sulphite mixtures and further from synthetic ethanol using non-yeast methylotrophic strains. Here, molasses distillers serve as a yeast growth stimulator, as a source of potassium, and optionally as a neutralizing agent (see Certificate No. 203 413 and No. 174 431). SUMMARY OF THE INVENTION The present invention relates to a process for the production of yeast products from methanol, wherein the yeast methylotrophic strains are cultured in a medium containing a medium alcoholic stillage in an amount of 0.2 to 0.3% by weight. 0.5 to 1% by weight, at a temperature of 30 to 40 ° C and a pH of 3.0 to 3.5.

As methylotrophic strains of yeast Candida can be used to dilute 11Bh, C. boidinii 1, Hansenula polymorpha, Pichia or mixtures thereof under sterile culture conditions, or acidotolerant methylotrophic yeast strains of Candida boidinii 2a Hansenula sp. 3 under non-sterile culture conditions.

Molasses stillage serves not only as a source of growth substances and mineral salts, but also for the growth of methylotrophic yeasts on media with methanol, but also shortens the cultivation time. Methylotrophic yeast in the presence of molasses stillage grows on medium with methanol immediately after ionoculation with maximum growth rate. When inoculumed on a new methanol-free medium without molasses stillage, methylotrophic yeasts show 2 to 6 hours of lag, almost without growth, depending on the size of the inoculum and aeration. The molasses stillage rooms increase the yeast biomass yield by more than 40%, which is the maximum yield of methylotrophic yeast relative to the consumed methanol. In the presence of molasses stillage, not only the yield of biomass but also the yield of all the specific products produced from the yeast biomass obtained can be increased. When enriching the medium with molasses stillage, the concentration of yeast cells over 40 g of dry biomass in 1 liter of fermentation fluid can be achieved in conventional agitators and air-conditioned fermenters using the inflow method of dosing small amounts of dofermentor, i.e. by 10 g in 1 liter of medium more than in the medium. with methanol. By using molasses stillage in methanol fermentations, as in the case of sulphite or. Alcohol blends, replace potassium content in the calcining portion of potassium ions added to the medium in the form of potassium phosphate or potassium bisulphate and thus save these raw materials.

The biomass obtained on methanol and molasses stillage is identical in terms of nutritional value of α-protein content to that of methanol itself, ie, it contains 60 to 62% of crude proteins essential to essential amino acids such as lysine, leucine, threonine, valine, and vitamins B2, and further comprises a maximum level of Cyspecific enzymes such as alcohol oxidase, formaldehyde- and formal dehydrogenase, di-hydroxyacetonsynthesis, further catalase, reduced nicotinadenine dinucleotide (NADH) areduced glutathione (GSH). In principle, there are two ways to cultivate. The molasses stillage can be added to the mineral medium either once at the beginning of the fermentation or gradually during the cultivation at a concentration of 0 to 1 to 0.3% by weight. Example 1 In a laboratory mechanically stirred fermenter with a total volume of 5 liters and a filling of 2.5 liters, a non-sterile single-culture cultivation of Candida boidinii 2 in mineral medium and methanol was performed. The composition of the mineral media was: 1 liter of water taps, 3 g of NH4 Cl; 7 g of KH 3 PO 4 · 0.5 g of Mg 80 · 7 · H 2 O; 0.1 g NaCl 0.1 g yeast extract and 10 µg biotin. At the beginning of the culture, 1% by weight of methanol was added to the culture medium. and molasses stillage in an amount of 0.25 wt. During the culture, the pH of the medium was kept constant at 3.0 by automatic dosing of 10% NaOH and the temperature was maintained at 30 ° C. Fermenter fermentation was 0.5 to 1.0 liters of air per hour, stirrer speed 600 per sec. 1 Concentrated molasses stillage contained potassium at a concentration of 11% by weight. and total conc. 5 wt. The concentration of yeast cells after inoculation of the medium was 0.2 g per liter in ferrogen. The addition of molasses stillage to the medium completely removed the lag, increased the growth rate of the cells in the early stages of growth, and increased the biomass yield relative to methanol from 38% to 45% by weight. C. boidinii 2 cells grow with a 4% biomass yield on medium without methanol.

For comparison, a further three cultures with a number of molasses were performed as shown in Table 1.

Table 1

Effect of molasses stillage on the biomass yield of C. boidinii 2 from methanol.

Molasses stillage,. dry wt. (g.l A) 0 1.0 2.5 3.0 Dry weight incr. cells (g.r1) 3.8 '4.2 4.5 4.6 Biomass yield of methanol (%) 38 42 45 46 Example 2 In a 4-volume, 75 liters mechanically stirred fermenter with 30 to 50 liters of mineral medium loaded as in Example 1, a non-sterile single-phase cultivation of Candida boidinii 2 was performed. Methanol was fed in a feed-in form of methanol-lye mixture (methanol and NaOH) preferably methanol-ammonia mixture (methanol and ammonia water) depending on the pH of the medium that was maintained automatically at a constant level of 3.0 to 3.2. Molasses stillage was dosed separately and sequentially during cultivation at 2.5 g per liter of media once every 24 hours. The air flow and agitator revolutions were controlled so that the dissolved oxygen concentration in the medium was between 10 and 60% saturation. The culture temperature was as in Example 1, i.e. 30 ° C. The amount of biomass obtained in the fermenter was 40 to 45 g of dry cells in 1 liter of medium. Example 3

The procedure was as in Example 2 except that the yeast strain Hansenula sp. 3 at 37-40 ° C

Claims (3)

EXAMPLE 4 In a 75-liter, mechanically stirred fermenter with a capacity of 30 to 50 liters of mineral medium composition As shown in Example δ. 1, a non-sterile continuous cultivation of Oandida boidinii 2 was performed, the medium's growth rate was in the range of D 0.05 h ”1 to 0.15 h” 3, preferably at D 0.1 h -1. 5% to 1% by weight and molasses stillage at a concentration of 0.25% by weight were fed to the fermentor together with the medium The pH of the medium and the culture temperature were maintained automatically as in Example 1. The yield of yeast biomass was 42 to 45 EXAMPLE 5 The procedure of Example 3 was followed except that methanol was dosed separately from the medium at a concentration of 25% by weight, depending on the dilution rate and the dissolved oxygen concentration in the medium that was maintained automatically. the pH of the medium was kept constant at pH 1 3.0 to 3.2 by automatic dosing with 50% NaOH, KOH or ammonia water. the difference, that was after at the cultivation temperature of 37 to 40 ° C. The yeast biomaea obtained by the method of the invention is rich in proteins, essential amino acids and vitamins and can be used and advantageously used as a valuable proteinaceous and vitaminic feed for livestock and as a source for specific enzymes such as alcohol oxidase, formaldehyde- and dehydrogenase, dihydroxyacetone synthases, catalase, nicotine adenine dinucleotide, adenosine triphosphate (ATP) and NASH, GSH, from methanol to a single, fed-batch, and continuous culture of yeast methylotrophs. <SUBJECT VT-1XIIZ A process for the production of yeast products from methanol, characterized in that the yeast methylotrophic strains are cultured in a medium containing molasses distillers in an amount of 0.2 to 0.3% by weight and methanol in an amount of 0.5 to 1% by weight at 30 ° C. to 40 ° C and pH 3.0 to 3.5 ·. .2. 12. The method of claim 1, wherein the methylotrophic strains of yeast are Oandida boidinii IIbh, C. boidinii 1, Hansenula polymorpha, Pichia, or mixtures thereof under sterile conditions, or acidotolerant methylotrophic yeast strains Canidida boidinii 2 and Hansenula sp. 3 under non-sterile culture conditions.