and/or steam
10 explosion with or without addition of acids resulting in a liquid fraction containing hemicellulosic sugars and a solid fraction containing cellulose and lignin. The liquid fraction ตามปกติ contains สารประกอบ that inhibit growth of microorganisms, สารประกอบ which are produced in the lignocellulosefractionation process. These สารประกอบ are degradation production of lignocellulose,such as lignin and sugars,
15 and ประกอบรวมด้วย สารประกอบฟีโนลิก, furan สารประกอบ (furfural and derivates thereof) and organic acids (mainly acetic acid, formic acid). Also the solid fraction containing cellulose and lignin contains inhibitory สารประกอบ, depending on the extent of washing. According to the การประดิษฐ์, the อาหารการเพาะเลี้ยง ประกอบรวมด้วย a liquid stream from fractionation step consisting hemicellulose sugars can be fed to
20 cultivation without enzymatic การไฮโดรไลซ์ of sugar oligomers, or alternatively hemicellulosestream containing sugar oligomers can be fed to enzymaticการไฮโดรไลซ์ to produce sugar monomers prior to be used in microbial cultivation. According to การประดิษฐ์, the solid cellulose-ligninfraction is fed to enzymatic treatment to dissolve cellulose and residual hemicellulose(not dissolved in the fractionation step) to sugar
25 monomersfor microbialoil production.
The ไลปิด ตามปกติ accumulate as intracellular lips within the จุลินทรีย์สะสมน้ำมัน (referred to as the first microbe), however the microbial ไลปิด may also be secreted or at least partly secreted to the fermentation broth from which it may be recovered.
30 Thus, in one embodiment, the method further ประกอบรวมด้วย a step of recovering the accumulated ไลปิด from said first microbe (จุลินทรีย์สะสมน้ำมัน). In อีกหนึ่งรุรุ,the ไลปิด is recovered from the fermentation broth. ไลปิด recovery may be carried out in various ways as discussed herein.
The incubation (cultivation) step (iii) may be performed as any suitable aerobic cultivation ที่รวมถึง batch, fed batch or continuous cultivation.
Where cultivation medium, the fermenter (เครื่องปฏิกรณ์ชีวภาพ) or systems connected to with
5 the fermenters has not been sterilized cultures of contaminating microbes may establish. It follows that where such contaminating microbes are not จุลินทรีย์สะสมน้ำมันs they may compete with the จุลินทรีย์สะสมน้ำมัน on the available substrate and thereby reduce the ไลปิด yield in the production.
10 These contaminating microbes (referred to a second microbe) are unwanted and should be avoided or suppressed in the system. By the introduction of a microbial inhibitor to which the จุลินทรีย์สะสมน้ำมัน is tolerant, the establishment of contaminating microbes (second microbe) avoided or suppressed in the system,
where the latter is sensitive or at least less tolerant to said microbial inhibitor.
15
Where the ไฮโดรไลเซตประเภทลิกโนเซลลูโลส has hot been sterilized, it will ตามปกติ contain one or more สปีชีส์ of จุลินทรีย์ที่ไม่ใช่แบบสะสมน้ำมันs, which are therefore unwanted in the cultivation and falls within the definition of the second microbe. Thus, in one embodiment said second microbe is a จุลินทรีย์ที่ไม่ใช่แบบสะสมน้ำมัน.
20
In one embodiment of the การประดิษฐ์นี้, the second (จุลินทรีย์ที่ไม่ใช่แบบสะสมน้ำมันs) is introduced or has established in the production system, e.g. the fermenter and thus will contaminate the• fermentation broth. Thus, in one embodiment of the
การประดิษฐ์, the fermentation broth further ประกอบรวมด้วย a second microbe, which is
25 intolerant to said at least one microbial growth inhibitor. Alternatively the second microbe is introduced during preparation for the cultivation, for example with the lignocellulosic hydrolysate. Accordingly, in a further embodiment, the second microbe is present in the cultivation medium provided in step (i) or contaminated the
fermentation broth at step (ii) or (iii) or present in the เครื่องปฏิกรณ์ชีวภาพ.
30
The object of the การประดิษฐ์นี้ is therefore to avoid that these contaminating microbes (referred to a second microbes) establish in the system. The is accomplished by introducing at least one microbial growth inhibitor to which the จุลินทรีย์สะสมน้ำมัน (first microbe) is tolerant or at least more tolerant and the
contaminating microbes (second microbe) is intolerant or at least less tolerant to than the first microbe.
It is evident from the above description that if the first microbe is "tolerant" to a
5 microbial growth inhibitor, it means that said first microbe (the oil producing microbe", is able to proliferate and/or produce oil even in the presence of said microbial growth inhibitor.