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Htp genomic engineering platform for improving fungal strains

A genome and genetic technology, applied in genetic engineering, fungi, viruses/phages, etc., can solve problems such as unfavorable, high viscosity, and affecting fungal growth

Pending Publication Date: 2020-01-21
ZYMERGEN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In some cases, due to the growth of mycelial fungi, the viscosity of the broth becomes high enough to significantly interfere with the dissolution of oxygen and nutrients, thereby adversely affecting the growth of the fungi and ultimately any desired product of interest. yield and yield

Method used

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  • Htp genomic engineering platform for improving fungal strains
  • Htp genomic engineering platform for improving fungal strains
  • Htp genomic engineering platform for improving fungal strains

Examples

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example

[0725] The following examples are given for the purpose of illustrating various embodiments of the invention and are not intended to limit the invention in any way. Those skilled in the art will recognize variations therein and other uses encompassed within the spirit of the invention as defined by the scope of the claims.

[0726] The following provides a brief navigation (ie Table 5) for the purpose of assisting the reader only. Nothing in this navigation is intended to limit the scope of the examples or explanations of this application. Table 5 - Navigation of the Examples Section

[0727]

example 1

[0728] Example 1: HTP genome engineering of filamentous fungi: generation and storage of filamentous fungal protoplasts

[0729] production of protoplasts

[0730] like Figure 20A As indicated in , 100 ml of complete medium was inoculated with 10 6 conidia / ml Aspergillus niger and grown overnight at 30°C at 150 rpm. After overnight growth, mycelia were harvested by filtering the culture through Miracloth. Subsequently, the mycelium was rinsed thoroughly with sterile water. For the experiments described in the Examples below, two strains of A. niger, A. niger strain 1015 and A. niger strain 11414, were used. Next, the collected and washed mycelia were subjected to enzymatic digestion by using VinoTaste Pro (VTP) enzyme mix. exist Figure 43 Characterization of each strain is depicted in .

[0731] For A. niger strain 1015, enzymatic digestion was performed by first preparing 50 ml of 60 mg / ml VTP-containing protoplast production buffer (1.2M magnesium sulfate, 50 mM pho...

example 2

[0734] Example 2: HTP Genome Engineering of Filamentous Fungi: An Alternative Approach to Protoplast Generation

[0735] like Figure 27 As indicated in , 500 ml of complete medium was inoculated with 10 6 conidia / ml Aspergillus niger and grown overnight at 30°C at 150 rpm. After overnight growth, mycelia were harvested by filtering the culture through Miracloth. Subsequently, the mycelium was rinsed thoroughly with sterile water. Next, the collected and washed mycelia were subjected to enzymatic digestion by using VinoTaste Pro (VTP) enzyme mix.

[0736] Enzymatic digestion was performed by first preparing 50 ml of 60 mg / ml VTP-containing protoplast production buffer (1.2M magnesium sulfate, 50 mM phosphate buffer, pH 5). After dissolving the VTP, the buffer was placed in a clean Oakridge tube and spun at 15,000 xg for 15 minutes. The sterile solution was then filtered after centrifugation. Once made, some of the collected mycelium was added to the VTP solution, and the...

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Abstract

The present disclosure provides an HTP genomic engineering platform for improving filamentous fungal cells that is computationally driven and integrates molecular biology, automation, and advanced machine learning protocols. This integrative platform utilizes a suite of HTP molecular tool sets to create HTP genetic design libraries, which are derived from, inter alia, scientific insight and iterative pattern recognition.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Application Serial No. 62 / 515,907, filed June 6, 2017, which is hereby incorporated by reference in its entirety for all purposes. technical field [0003] The present invention relates to automated fungal genome engineering. The disclosed automated genome engineering platform requires genetic manipulation of filamentous fungi to generate fungal production strains and facilitate their purification. The resulting fungal production strains are well suited for growth in submerged cultures, eg, for large-scale production of products of interest (eg, antibiotics, metabolites, proteins, etc.) for commercial applications. [0004] Statement Regarding the Sequence Listing [0005] The Sequence Listing associated with this application is provided in text format instead of a paper copy and is hereby incorporated by reference into this specification. The name of the text...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1058C12N15/1079C12N2310/20C12N1/14C12N9/22C12N15/11C12N15/80C12N2800/80
Inventor V·松斯皮拉J·弗雷德隆德H·阿卜杜拉P·博卡齐S·普斯特S·D·L·A·克莱托B·柴金德D·沃恩K·S·布鲁诺P·韦斯特福尔E·谢夫奇克K·罗特席尔德-曼奇内里A·M·方三世
Owner ZYMERGEN INC
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